Biological Markers in Psychiatry and Neurology: Proceedings of a Conference Held at the Ochsner Clinic, New Orleans, on May 8-10, 1981

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ABBREVIATIONS

AA = alcohol preferring (rats)

AANB = α-amino-n-butyric acid

ACh = acetylcholine

ACTH = corticotropin

ADH = alcohol dehydrogenase

ADTN = 6,7-dihydroxy-2-aminotetralin

AEP = auditory-evoked potential

AER = average evoked response

A-I = Activation-Inhibition Scale

AIR = average impairment rating

AMP = adenosine monophosphate

AMPT = α-methyl-p-tyrosine

ANA = alcohol avoiding (rats)

ANOVA = analysis of variance

ANS = autonomic nervous system

ATP = adenosine triphosphate

ATPase = adenosine triphosphatase

BAC = blood alcohol concentration

BEAM = brain electrical activity mapping

BP = bipolar (depression)

BPRS = Brief Psychiatric Rating Scale

CaM = calmodulin

Ch = choline

CNS = central nervous system

CNV = contingent negative variation

COMT = catechol O-methyltransferase

C/P = cholesterol/lipid-P ratio

CPB = competitive protein binding (assay)

CPT = continuous performance task

CRU = Clinical Research Unit (of WPIC)

CSF = cerebrospinal fluid

CSU = Clinical Studies Unit (U. of Mich.)

CT = computerized (axial) tomography

CV = coefficient of variation

DA = dopamine

DBH = dopamine β-hydroxylase

DF = discriminant function

DHEC = dihydroergocryptine

DNA = deoxyribonucleic acid

DOPA = dihydroxyphenylalanine

DOPAC = dihydroxyphenylacetic acid

DSD = depression spectrum disease

DSM-III = Diagnostic and Statistical

Manual of Mental Disorders – 3rd Ed.

DST = dexamethasone suppression test

D.Z. = dizygotic (twins)

ECT = electroconvulsive (shock) therapy

ED = endogenous depression

EDTA = ethylenediamine tetraacetic acid

EEG = electroencephalogram

EMG = electromyogram

EOG = electrooculograph or electrooculogram

EP = evoked potential

EPI = epinephrine

ERP = event-related potential (s)

FA = fusaric acid

FFT = fast Fourier transform

FHN = family history negative

FHP = family history positive

FH-RDC = family history-Research Diagnostic Criteria

FPDD = familial pure depressive disease

FSH = follicle stimulating hormone

FTG = fastigial tegmental gigantocellular (field)

FTI = free thyroid index

GABA = γ-aminobutyric acid

GFAP = glial fibrillary acidic protein

GGNRM = uniform normal random number generator

GGT = γ-glutamyl transferase

GGTP = γ-glutamyl transpeptidase

GH = growth hormone

Gpp (NH)p = guanylimidophosphate

GSR = galvanic skin response

GTP = guanosine triphosphate

α2H = α2 receptor-high affinity

HAT = high accuracy tracking

HDRS = Hamilton Depression Rating Scale

hGH = human growth hormone

5-HIAA = 5-hydroxyindole acetic acid

HLA = human leukocyte antigen

HPA = hypothalamic-pituitary-adrenocortical (system)

HPL = hypothalamopineal pathway lesion

HPLC = high performance liquid chromatography

HR = heart rate

5-HT = serotonin

HVA = homovanillic acid

IMSL = International Mathematics Science Library

IR = infrared (recorder)

ITT = insulin tolerance test

α2L = α2 receptor-low affinity

LAT = low accuracy tracking

LC = locus coeruleus

LDH = lactate dehydrogenase

LH = luteinizing hormone

Li = lithium (cation)

LR = lithium ratio

MANOVA = multivariate ANOVA

MAO = monoamine oxidase

MCV = mean corpuscular volume

MHPG = 3-methoxy-4-hydroxyphenylglycol

MIDAS = Michigan Interactive Data Analysis System

MIF-l = prolyl-leucy-glycinamide

MMPI = Minnesota Multiphasic Personality Inventory

MSH = melanocyte stimulating hormone

M.Z. = monozygotic (twins)

Na-dep. = sodium-dependent

ND = non-endogenous (neurotic) depression

NE = norepinephrine

NHSI = New Haven Schizophrenic Index NIAD = National Institute of Allergy & Infectious Diseases

NIMH = National Institute of Mental Health (Rating Scale)

NIMH-D = NIMH-depressed group

NIMH-N = NIMN-normal group

NT = non-target (s)

OR = orienting responses

PAC = p-aminoclonidine

PAL = pituitary-adrenal axis lesion

PC = phosphatidylcholine

PCPA = p-chlorophenylalanine

PE = phosphatidylethanolamine

PEA = phenylethylamine

PEG = pneumoencephalography

PF = personality factors

PGE1 = prostaglandin E1

POMS = Profile of Mood States

PRBS = pseudorandom binary sequence

PRP = platelet-rich plasma

PS = phosphatidylserine

PSE = Present State Examination

PTC = phenylthiocarbamide

PV = predictive value (of a positive test)

R = receptor (s)

RAS = reticular activating system

RBC = erythrocytes

RDC = Research Diagnostic Criteria

REM = rapid eye movement (sleep)

RLMA = REM latency minus awake

RT = reaction time

SADS = Schedule for Affective Disorders and Schizophrenia

SADS-L = SADS-Lifetime Version

SC = skin conductance

SCL = SC level

SCR = SC response

SDD = sporadic depressive disease

SDS = sodium dodecyl sulfate

SGOT = serum glutamic-oxaloacetic transaminase

SGPT = serum glutamic-pyruvic transaminase

SHAS = Subjective High Assessment Scale

SP = sphingomyelin

SPEM = smooth pursuit eye movement

SPM = significance probability maps

SPV = (reconstructed) slow phase velocity

T = target (s)

T3 = triiodothyronine

T4 = thyroxine

TEMED = N, N, N′, N′-tetramethylethylenediamine

TRH = thyrotropin-releasing hormone

TSH = thyroid-stimulating hormone

UFC = urinary free cortisol

UP = unipolar

VEP = visual-evoked potential

VMA = 3-methoxy-4-hydroxymandelic acid

VOR = vestibulo-ocular reflex

WAIS = Wechsler Adult Intelligence Scale

WHO = World Health Organization

WISC = Wechsler Intelligence Scale for Children

WPIC = Western Psychiatric Institute & Clinic (University of Pittsburgh)

YOH = yohimbine

I

NEUROCHEMICAL MARKERS

HIGH RISK STUDIES IN AFFECTIVE DISORDERS

D.L. Dunner,     Department of Psychiatry and Behavioral Sciences, University of Washington, and Harborview Medical Center, Seattle, Washington, USA

ABSTRACT

Several biological markers have been studied in depressed patients. Whereas some of these studies support an association of a marker with the depressed state, no trait markers for depression have as yet been developed which would provide a biological basis for high risk studies.

KEYWORDS

Affective disorders

High Risk

biological markers

INTRODUCTION

The purpose of this paper is to review research regarding children at risk to develop depression. Research in depression changed to a psychobiological focus in the 1970’s. The rationale for this emphasis related to refinements in clinical classification of affective illness, classifications which were supported by studies showing an increased incidence of psychosis and mania among relatives of bipolar as compared with relatives of unipolar patients (Leonhard, Korff, and Schultz, 1962). The recognition that some depressions may have a genetic etiology supported research in depression which tried to reveal the biology of these illnesses. Further support for such studies came from psychopharmacology, as the use of antidepressant medication in the treatment of depression increased.

In this paper we will review the attempts to provide a biological definition of depression. Strategies for high risk studies will be discussed.

Classification of Depression

It is logical to assume that biological factors will be associated with some but not all depressions. A classification of depression which separated biological affective states from other syndromes would enhance the attempt to identify biological factors related to depression, or even uncover the etiology of a depressive subtype. The unipolar-bipolar classification seemingly fits such a classification. This dichotomy initially separated patients on clinical grounds (presence or absence of a history of mania) and was supported by familial data. Subsequent family studies clearly supported the homogeneity of bipolar illness. Mania among relatives of depressed probands is largely confined to families of bipolar probands (Perris, 1966; Winokur, Clayton and Reich, 1969; Dunner, Gershon, and Goodwin, 1976). Initial data supporting the separate classification of patients with histories of mania (Bipolar I) and, later, of hypomania (Bipolar II) were derived from family, pharmacological, and biological studies (Fieve and Dunner, 1975). These data suggest that Bipolar I patients represent a fairly homogeneous population with regard to the above mentioned factors. However, an etiology (or etiologies) for Bipolar I illness has not been demonstrated. In contrast, unipolar disorder is clinically heterogeneous, and attempts to validate clinical subdivisions of unipolar illness have been supported in the main by genetic data only (Winokur and co-workers, 1971).

The current nomenclature (DSM III) acknowledges the clinical validity of bipolar depression, although the bipolar affective disorder of DSM III has greater clinical heterogeneity than prior classifications. Unipolar disorders have been aggregated in DSM III under the term Major Depressive Disorder.

The adoption of DSM III has occurred, however, without a supporting base of data bridging these new subclassifications with prior subgroups. Many studies of depression prior to DSM III utilized the criteria of Feighner and co-workers (1972) of primary affective disorder. Patients who had primary affective disorder were subsequently separated into bipolar and unipolar subtypes. Cases of secondary depression, usually unipolar but at times bipolar, were excluded from these studies. Hopefully, future studies will focus on the relationship of DSM III subtypes to those proposed earlier, and data will be generated to support the DSM III classification. In lieu of these data, we will in this paper by necessity report on the older studies which employed the criteria of Feighner et al (1972) for primary affective disorder.

State versus Trait Markers

Factors which are found in higher frequency among depressives than controls may serve to identify an ill population. These factors may be clinical (psychological profiles or sleep patterns), biological (concentrations of catecholamine metabolites in the urine), physiological (cortical evoked response), pharmacological (clinical response to medication or biological response to medication), and genetic (family history). Factors which are present only during an episode of illness are considered state markers. Such factors may be of benefit in depression in establishing the diagnosis or in evaluating response to treatment. The dexamethasone suppression test (DST) (Carroll, 1980) is an example of a valuable state marker. Failure to suppress serum coritisol after a dose of dexamethasone occurs in some patients with depression. A limited number of medical conditions may also be associated with dexamethasone non-suppression. Thus far, no other group of psychiatric patients has shown non-suppression with the DST. Therefore, this test has specificity for the ill state. Non-suppression to DST reverts to normal as the depression improves. The DST can be used to diagnose depression and can also be used to determine when patients have recovered. The DST is state (illness) dependent, since it apparently is not positive in recovered depressives.

A trait marker, in contrast would be positive whether the person had the symptoms of the clinical syndrome or not. An example of a trait marker is the finding of S hemoglobin among carriers of sickle cell trait. In contrast to state markers, research directed at determining trait markers occurs in individuals who have recovered from depression or are likely to become depressed. The factor should be found in recovered depressives but not in controls. If the factor is found in a familial form of depression, then the factor should also be found in greater frequency among relatives of patients than in controls.

Thus far, no trait markers relevant to depression have been documented. In this paper some of the pertinent state markers will be discussed. It should be stressed, however, that trait markers would be necessary to validate findings of high risk studies.

The Catecholamine Theory: MHPG

The catecholamine hypothesis of affective disorder was the major theory in the 1960’s regarding biological factors in depression (Schildkraut, 1965). This hypothesis suggested that some depressions may be associated with relative deficiencies in norepinephine in certain areas of the central nervous system. Testing this hypothesis necessitated a series of studies of catecholamines and metabolites of catecholamines in affectively ill patients. Such studies were hampered by inadequate laboratory methodology. Although initial efforts were directed at catecholamine (and particularly norepinephrine metabolism), it soon became evident that other neurotransmitters (particularly dopamine and serotonin) were of interest and the theory was expanded to a biogenic amine theory of depression (in order to include indoleamines as well as catecholamines). Various strategies emerged, including the measurement of concentrations of amines and/or their metabolites in urine, cerebrospinal fluid, and blood of patients, assessing the clinical effect of administration of medications affecting central nervous system metabolism of amines, and measuring the activity of enzymes related to amine synthesis and degredation. These strategies were largely state related and generally preceded a recognition that familial factors may be important in refining the classification of affective illness.

There are several findings which support a state-dependent relationship between patients with affective disorders (at times separated into subtypes) and biological markers related to biogenic amines. The current catecholamine metabolite of interest is MHPG (3-methoxy,4-hydroxyphenylglycol), a norepinephrine metabolite usually measured in collections of urine. It is thought that a considerable portion of the urinary concentration of MHPG derives from the central nervous system rather than from peripheral noradrenergic nerves. When studied in cycling patients, urinary MHPG is increased during mania as compared with the same patient during depression. In general, MHPG is lowest among patients with bipolar depression and higher among patients with unipolar depression (Shildkraut, 1978). It has been suggested that (among unipolar depressives) higher excretion of MHPG predicts an antidepressant response to amitriptyline whereas lower excretion of MHPG predicts response to imipramine (Beckmann and Goodwin, 1975). This finding is relative to the underlying hypothesis since imipramine has a greater effect on noradrenergic neuronal reuptake than does amitriptyline, which has a reuptake effect more selective for serotonin.

A factor which parallels the MHPG finding is psychomotor activity. Bipolar depressives usually evidence psychomotor retardation whereas unipolar depressives tend more frequently to be agitated and anxious (Beigel and Murphy, 1971; Dunner, Dwyer, and Fieve, 1976). Furthermore, amitriptyline is clinically preferred to imipramine in patients whose depressions are anxious, sleepless, and agitated. Imipramine, which is less sedating and more activating, is clinically preferred for depressions characterized by anergy.

Although there have been attempts to assess the influence of activity on MHPG excretion, it is not entirely clear that the entire relationship of urinary MHPG to depression is not a mere reflection of patient motor activity, with lower values found among psychomotor retarded (bipolar) patients and higher values found among agitated-anxious (unipolar) patients.

The demonstration that high MHPG excreting amitriptyline responders are also imipramine non-responders and that lower MHPG excreting imipramine responders are amitriptyline non-responders is seemingly crucial in establishing MHPG as a state marker for depression rather than an indicator of psychomotor activity. Such a study has not been definitely reported.

Biogenic Amines: Enzymes

Another series of studies related to the biogenic amine hypothesis of mood disorders involved the activities of enzymes involved in the synthesis and metabolism of amines. The activities of these enzymes were measured in convenient peripheral tissues (plasma, erythrocytes, platelets) and were generally studied in patients who were both symptomatic and euthymic. Enzyme activity may be an ideal trait marker. Genetic deviation which results in the production of abnormal enzyme protein can lead to absent enzyme activity. The activity of Dopamine Beta Hydroxylase (DBH) in familial dysautonomia is a model for such studies. Thus, these enzymes have also been studied in relatives of patients.

The major studies in affective disorder patients relate to monoamine oxidase (MAO) and catecholamine-O-methyltransferase (COMT). COMT activity was initially found to be reduced in women with depression, particularly unipolar depression. A subsequent study noted increased COMT among depressives as compared with controls, and later the initial findings could not be replicated (Cohn, Dunner, and Axelrod 1970; Gershon and Jonas, 1975; Dunner and co-workers, 1977). The MAO studies revealed lower mean activity in bipolar as compared with unipolar patients. Subsequent studies failed to replicate this finding (Murphy and Weiss, 1972; Edwards and co-workers, 1978; Fieve and co-workers, 1980). Gershon (1978) replicated the finding among probands but failed to demonstrate that affectively ill relatives of low-MAO probands themselves had low MAO activity. The Gershon (1978) study serves as a model study for research into biological markers combining a genetic model strategy. They noted that a true trait marker should be present not only among ill patients but also among the ill relatives of such patients.

The early positive findings of affective illness and COMT and MAO should have been interpreted with greater caution. What was demonstrated was a significant difference in the mean activity of these enzymes between patients and controls. The range of individual values, however, overlapped. As larger series were reported, the initial findings could not be replicated. The lack of caution in interpreting these studies was probably related to a state rather than a trait model of investigation. Investigations prior to enzyme studies were based on showing differences between acutely depressed patients and controls. Abnormal enzymes often result in no activity, and absent activity was not demonstrated in any affectively ill patient in studies of COMT, MAO, or DBH.

Biogenic Amines: Precursor Loading

Biogenic amine precursors have been administered to affectively ill patients with interesting results. L-DOPA, a precursor of norepinephine and dopamine, was shown to have antidepressant effects among retarded depressives (Goodwin and co-workers, 1971). Subsequent review of the data revealed that Bipolar I depressed patients became manic-like or psychotic when given L-DOPA. An antidepressant response was shown with L-DOPA administration to Bipolar II depressed patients, whereas unipolar depressives showed no response to L-DOPA (Gershon and co-workers, 1971). Similarly, L-tryptophan, a precursor of serotonin, was given to depressed unipolar patients with no response, although some Bipolar patients (I and II) showed antidepressant responses to L-tryptophan administration (Farkas, Dunner, and Fieve, 1976). These data support the hypothesis of a deficiency of biogenic amines in bipolar depression. However, attempts to treat mania with inhibitors of biogenic amine synthesis have, in general, not been successful (Bunney and co-workers, 1971). The precursor load strategy has been employed only with ill patients. The administration of such compounds to patients with a history of affective disorder, or their relatives, would be of interest in determining whether response to such compounds is a trait marker for depression.

Cholinergic Models

Sleep patterns show characteristic alterations in depressed patients, who demonstrate a shortened period from sleep onset to the initial rapid eye movement (REM) sleep stage as compared with controls (Kupfer and co-workers, 1978). This finding seems to be independent of depressive subtypes and apparently is state dependent. Recently, Sitaram and co-workers (1980) administered an acetylcholine agonist (arecholine) to euthymic patients who had a history of affective disorders. These patients had a different pattern of response with arecholine regarding REM latency as compared with controls. Their data suggested cholinergic supersensitivity in some affective disorders. Further studies with arecholine may be of interest in establishing this test as a possible trait marker for depression.

The Lithium Pump

Lithium salts are effective in the treatment of bipolar illness (acute mania, acute depression, maintenance treatment for recurrent mania and depression) and unipolar illness (maintenance treatment for recurrent depression) (Dunner and Fieve, 1978). The maintenance effect of lithium is remarkable, and an understanding of the pharmacology and mechanism of action of lithium may be relevant to an understanding of the biology of affective illness.

Although lithium works within cells, the dose is commonly regulated through measurement of lithium concentration in plasma. Erythrocyte lithium concentration is about half the plasma concentration. Recent studies suggest that there is an active process for removal of lithium from cells, and that the rate constant for this process can be measured. Meltzer and co-workers (1976) express this rate constant for lithium efflux as Ko.

Ko has been studied in patients with affective disorder and controls and no differences have been shown (Dunner, Meltzer, and Fieve, 1978). There is general consensus that Ko is under some kind of genetic regulation since values for Ko within families show a higher correlation than values for non-related individuals. However, a large study of Ko failed to demonstrate that relatives with a history of affective disorder had different Ko values than controls, although there is lack of complete agreement from various centers (Dunner, Meltzer, and Fieve, 1979; Pandey and co-workers, 1979). In summary, Ko does not seem to be a suitable marker for depression state or trait.

Dexamethasone Suppression

Another state marker of depression is the finding of continuous elevations of serum cortisol. Dexamethasone administration results in a suppression of serum cortisol, and failure to show such suppression is found in certain disease states. Studies by Carroll (1980) clearly show that approximately half of a population of depressed patients will fail to show suppression of serum cortisol after dexamethasone administration-the so-called DST test. If one excludes medical causes of non-suppression, a positive test is an almost certain indication of the depressed state. Patients who are non-suppressors will become suppressors as their depression remits. Thus, the DST is not a trait marker, but is quite valuable as a state marker of depression, particularly for severe endogenous depression. The DST may also be helpful in characterizing depressed populations for other studies.

Psychological Testing

Psychological testing, including tests for intelligence, personality, and memory, have not been widely studied in patients with well characterized affective disorders. Liebowitz and co-workers (1979) administered the Maudsley Personality Inventory to a group of outpatients and found higher ratings for extraversion and lower ratings for neuroticism among bipolars as compared to unipolare. However, the findings largely disappeared when patients with minimal depressive symptoms were excluded and the analysis was performed on only strictly defined euthymic patients. Thus, some stable personality variables which may be of use as trait markers may be state dependent in their measurement.

Cohler and co-workers (1977) found that children of individuals with psychotic depression had lower scores on the Wechsler Intelligence Scale for Children (WISC) than children of well parents. Further studies of the WISC in children of bipolar and unipolar patients may be of interest, and it may be possible to use subscales of the WISC as trait markers.

High Risk Studies

In summary of the above, there have been several research efforts to develop markers for depression. Most of the putative markers thus far identified are state dependent. Research involving sleep, arecholine, the DST, and the WISC may be of particular importance in leading toward a trait marker. However, at the present time there is no well developed trait marker for depression.

High risk studies of depression are predicated on the illness (or subtype) being heritable and on a reliable trait marker which can identify those offspring who are likely to become ill in contrast to their siblings who will remain well. One goal of such studies is to study the environmental aspects of becoming ill. Thus, the high risk model is essentially a genetic model wherein one studies psychosocial, environmental, or other non-genetic factors involved in the development of illness.

Kuyler and co-workers (1980) reviewed some factors which seemed to be associated with psychopathology among offspring of a predominately bipolar population. Children with one ill parent had a lower rate of illness than if both parents were affectively ill. Divorce or prolonged separation of parents also seemed to result in an increase in psychopathology among the children. Another factor was age (older children were more likely to be ill than younger children). Age of onset of parental illness did not seem to be a factor in their study, in which only 4 of 49 children had a depressive disorder.

McKnew and co-workers (1979) studied 30 children of affectively ill parents and 9 met criteria for depression over a 4 month period. Since considerably less than half of adult relatives of bipolar patients have affective disorder (Dunner, Go, and Fieve, 1980), their high incidence of depression in children may be a result of applying diagnostic criteria which define too broad a sample of affective disorder. Diagnostic criteria for depression in childhood are much more problematic than in adults. Furthermore, the behavioral disturbance in childhood which is the antecedent of adult depression may or may not be a manifest depression.

Kuyler and co-workers (1980) outlined a procedure for high risk studies, involving studying both children of patients and children of controls. Controls could be selected from cousins of these children, providing some additional genetic data, as well as from children of patients with chronic medical illnesses. The ability to diagnose children reliably is critical, particularly if there is no trait marker to otherwise identify children with a high likelihood of manifesting adult depression. The children should be followed over time with attention paid to both the early development of symptoms as well as environmental influences. If those children who will become ill as adults can be reliably identified, therapeutic interventions could be made.

Key elements of the above involve a large and rigorously defined adult population, a trait marker, and a reliable and valid assessment instrument for the children. Hopefully further work in biological markers associated with affective illness will lead to the identification of suitable trait markers.

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DISCUSSION

Dr. Zubin wondered what we mean by the term markers. Dr. Dunner discussed state and trait markers; he suggested that the markers discussed up to this point had been used to identify individuals vulnerable to some pathology, but that not all markers are concerned with leads to disorders; some may be concerned with protection from disorders.

BIOLOGICAL MARKERS OF SEPARATION INDUCED DEPRESSION IN PRIMATE MODELS

W.T. McKinney, E.C. Moran and G.W. Kraemer,     Department of Psychiatry, Wisconsin Psychiatric Research Institute, and Primate Laboratory, University of Wisconsin-Madison, Madison, Wisconsin, USA

ABSTRACT

This chapter reviews biological markers of separation induced depression in nonhuman primates and other animals. Disruption of attachment bonds, whether in humans or nonhumans, has by now been established as a very stressful event. The behavioral effects of either maternal separation or peer separation are striking and resemble the depressions shown by some humans undergoing major separations.In addition to the behavioral effects of separation, there are a variety of neurobiological consequences including effects on sleep, heart rate, body temperature, steroids, brain self-stimulation rates, brain serotonin levels, catecholamine synthesizing enzymes in the adrenal gland, and CSF norepinephrine, 5-hydroxyindoleacetic acid, homovanillic acid. These studies are summarized in this chapter.The separation response in monkeys and several other animals is influenced by a variety of pharmacological manipulations. The drugs which have been tried thus far and their main effects are dicussed in the final section of this chapter.

KEYWORDS

Separation

monkeys

biological markers

behavior

drugs

depression

INTRODUCTION

This chapter is divided into three sections. The first section focuses on a review of the separation syndromes in nonhuman primates with some mention of separation effects in other species. The second section focuses on separation as an independent variable and reviews a variety of dependent neurobiological variables that have been studied in relationship to separation, i.e., how separation influences a variety of neurobiological systems. In the third and last section, we deal with various neurobiological influences on the separation reaction; that is, what is known about how different pharmacological manipulations affect the response to separation in experimental animals.

SECTION I REVIEW OF SEPARATION SYNDROMES IN MONKEYS

Disruption of attachment bonds, whether in humans or nonhumans, has by now been established as a very stressful event. It is likely that humans and many animal species are in their most stable condition when they have developed secure social attachment systems. Disruption of such attachment systems leads to serious sequelae including in some instances clinically significant depressions. Many behavioral, developmental, and neurobiological variables undoubtedly influence the response to separation. Delineating the exact nature of the influence of each of these variables and how they might interact with each other has been a difficult task in research with humans, and for this reason animal models have been increasingly used for a more systematic study of the effects of separation.

The first reported experimental separation of monkey infants from their mothers occurred in the 1960’s. Pigtail macaque (M. nemestrina) infants were separated from their mothers at ages of five and seven months for short periods of time and then reunited with their own or another mother (Jensen and Tolman, 1962). Also in 1962, Seay, Hansen, and Harlow separated rhesus macaque (M. mulatta) infants from their mothers at about the same age but for longer periods of time. Both groups reported these separations as being traumatic events for the infants resulting in increased arousal and increased distress vocalization. The behavior patterns seen in human children who were separated from their parents was compared with those observed in infant monkeys that had been separated from their mothers. These behaviors were divided into two categories labeled protest and despair and deemed similar to the responses reported by Spitz (1946), Bowlby (1960), and the Robertsons (1971) who originally described these phases in human children who had been separated from their families. This biphasic protest and despair reaction is by now well described in the primate literature.

The protest stage for monkey infants generally consists of a disoriented increase in activity accompanied by loud and repeated screeching. If the mother is visible there are frantic attempts to reach her. This stage generally lasts 24-36 hours followed by the despair stage which consists of decreased activity, food and water intake, and vocalizations with generalized social withdrawal. In some instances, this general decline is so severe that it results in the animal’s death. This can certainly occur if fluid and electrolyte balance is not maintained but sometimes happens even in the face of external maintenance of these vital physiological functions.

Research over the years has established that there are a number of different factors which affect the response to maternal separation. Rosenblum and Kaufman (1968) reported a study in which species of the experimental subjects seemed to be an important variable. When they separated bonnet macaque (M. radiata) infants from their mothers, only a mild protest reaction and almost no despair response were noted. This is a very different response from that shown by both rhesus and pigtail infants and may be explained by the nature of the social patterns of a particular species. Bonnet infants spend much more time interacting with other members of the group and the general behavior of group members is more permissive and responsive to infants than other species. There is less dyadic bonding between a biological mother and her infant and more group parenting in the case of the bonnet macaque. By contrast, pigtail macaques form strong dyadic social bonds and when the biological mother is removed from the group, there is minimal substitute mothering and the pigtail infants undergo a profound depressive- type response.

Age and social situations can also affect the response to maternal separation. When Suomi, Collins, and Harlow (1973) separated rhesus infants from their mothers at 60, 90, and 120 days of age, they found that the monkeys exhibited a typical protest and despair response, and that the infants that were separated at 90 days reacted much more adversely than infants separated at other ages. They further reported that infants that were housed in single cage units during the separation period showed more severe depressive behaviors than infants which were housed with peers. These differences lasted and even became worse by six months of age.

Sex and preexisting behavior patterns have also been indicated as contributing factors in the response to maternal separation. Hinde and his colleagues (1966, 1972, 1974) separated rhesus infants from their mothers or vice versa for short periods of time. They also reported the existence of protest and despair stages and presented data to indicate that preseparation behavior is predictive of behavior which occurs after separation and that sex of the infants may also be a factor in response to maternal separation with male infants responding more severely than female infants. This same group has also found that short-term maternal separation early in development has long-term effects some two to three years later.

A number of maternal separation studies have been done which also measured biological parameters and these will be reviewed in Section II of this chapter.

Another aspect of separation that is important to understand from a strictly behavioral standpoint is the peer separation model. It too, is based on the fact that rhesus monkeys develop sophisticated social bonds. While in no way denigrating the unique bonds of mother and infant in some ways the peer relationship has some special advantages. The mother- infant affectional system is obfuscated by the fact that the infant is totally dependent on the mother for food and protection as well as social contact. In addition, it is by its very nature a social system which is rapidly changing and relatively short-lived. The peer separation paradigm provides a more stable and longer lasting system in which to do both behavioral and neurobiological studies. The protest and despair patterns seen as a result of separation from peers are similar to those seen after maternal separation. The development of another animal model of depression utilizing peer separation does not lessen the value of the mother- infant model but rather offers another opportunity to study parameters that cannot be investigated in the mother-infant paradigm.

Suomi, Harlow, and Domek (1970) and Bowden and McKinney (1972) have suggested peer separation as a viable animal model of depression.

Rhesus infants that had been separated from their mothers at birth and housed with peers at 15 days of age showed strong protest and despair responses that were substantially the same on the twentieth separation as during the first one. Subsequent research indicates that frequency of separations is not the decisive factor in influencing the nature of the despair response to peer separation.

Suomi and Harlow (1972) and Suomi (1976) separated peer-reared rhesus monkeys and housed some in single cages and some in vertical chambers that allowed no visual or physical contact with the environment or other animals. Both groups of infants responded severely to separation but the group housed in the vertical chamber responded more severely than the group housed in single cages. Thus it appears that the environment influences the despair response to separation.

Suomi (1976) found that the behavioral profiles of infant monkeys before separation helped predict their reaction to subsequent separation. Specifically, infants that had been reared with their mothers for three months before being put into peer groups, responded less severely to separation than infants that had been peer-reared from birth or shortly thereafter. Kraemer (1978) reported that peer-reared subjects showed a more adverse response to separation from their peers than mother-reared monkeys. Therefore, it appears that the preseparation rearing condition as well as the housing environment during separation influences the rhesus infant’s response to separation.

Bowden and McKinney (1972) separated pairs of juvenile rhesus monkeys for two weeks after they had been housed with each other for eight months. The study reported signs of mild protest but no despair. A later study by McKinney, Suomi, and Harlow (1972) reported similar results when juvenile monkeys (three years old) who had been reared together since infancy were repeatedly separated for four 2 week periods. Thus age is also an important variable in influencing the nature and form of the separation response.

These studies of separation in juvenile peers, as a whole, suggest a number of different explanations. One possibility is that adolescent monkeys do not have the intense emotional response to separation from peers that younger monkeys do. It may also be that these animals showed their response to separation in other ways but were equally disturbed. Another possiblity is that the animals in these studies simply were not together long enough to form strong attachment bonds that must be present before the effects of its loss can be manifested. This is further supported by a 1975 study by Suomi and colleagues (1975) who separated five-year old rhesus monkeys from their nuclear family. The nuclear family is a highly enriched living situation by laboratory standards. The monkeys were removed from the nuclear family and then housed with familiar peers, strangers, or alone. In each of these situations, they displayed qualitatively different responses. All of the animals were disturbed, but those that were housed with familiar monkeys or strangers returned to their preseparation behaviors in a short time. However, the monkeys that were housed alone showed the classic protest and despair response to separation. This study indicates that the traditional despair response can occur in older monkeys undergoing separation as well as in infants.

SECTION II STUDIES OF NEUROBIOLOGICAL VARIABLES AS INFLUENCED BY SEPARATION

Reite and associates (1976, 1977, 1978a, 1978b, 1978c) have done a series of studies of pigtail macaque infants undergoing maternal separation. Using totally implanted multichannel biotelemetry systems, they have studied body temperature, heart rate, and sleep physiology of the infants before, during, and after separation from their mothers. From a behavioral standpoint, they have found that attachment bonds are as central to the development of monkeys as they are to people, and are crucial for the maintenance of family and social structure and for the ability of the individual to relate to such a structure. The disruption of these bonds has been found to be linked with the development of serious psychopathology and the alteration of certain physiological processes.

In one such study of ten group living pigtail macaques, they found significant increases in the infant’s heart rate and body temperature immediately after maternal separation. These changes were most pronounced very early in separation and tended to diminish as the separation continued. Beginning with the first night both the heart rate and body temperature showed marked decreases from baseline levels and the behavioral patterns became more depressive-like. During reunion both the heart rate and body temperature returned to normal although some infants showed the lower heart rate well into the reunion. Significant sleep changes have also been reported to occur during maternal separation. The reported changes have included increased sleep latency, more frequent arousals, less total sleep, and a disruption of REM