Principles of Development

Introduction
How does a single fertilized egg develop into an embryo and then into a baby and eventually an adult? o Fundamental principles are common to all developmental processes observed in multicellular organisms! Developmental biology is an interdisciplinary field biochemistry, cell biology, genetics and evolution

Shared Developmental Processes

Five essential cellular processes in development: o Cell division (proliferation) o Programmed cell death (apoptosis) o Cell movement and differential expansion o Differential gene expression and cell differentiation o Cell-cell interactions

Cell Proliferation
For an individual to develop from an undifferentiated mass of cells, its cells have to proliferate (divide and make more cells) o Location, timing, and extent of cell divisions are tightly controlled by interacting layers of regulation recall MPF and signaling from other cells Most cells stop proliferating at maturity o are some specialized, undifferentiated cells that continue proliferating throughout the organisms life In animals = stem cells

Programmed Cell Death (Apoptosis)

Carefully regulated aspect of normal development Abnormal apoptosis either too much or too little can lead to disease or deformation

Cell Movement
In addition to dividing, many animal cells have to move for normal development to occur. o In animals, during gastrulation cells in different parts of early embryo rearrange themselves into three distinctive types of embryonic tissues which later form specific organs

Cell Differentiation
During development, most cells must undergo differentiation in order to become a specialized type of cell involves changes in gene expression o Differentiation is a progressive, step-by-step process. Cells are initially determined (committed) to a specific developmental pathway and later become differentiated

Cell Differentiation
Animal embryonic cells in early stages of development are totipotent capable of becoming ANY cell o Animal cells are unable to de-differentiate Animal stem cells do not become specialized adult cells, but instead remain undifferentiated o Stem cells: retain the ability to divide and give rise to an array of specialized cell types

Cell-Cell Interactions
During development, the most important cell-cell interactions involve sending and receiving signals! o Cell-cell signals change patterns of gene expression o Cell-cell signals are essential for changing cell activity during development

Developmental Biology is Interdisciplinary

Recall from Chapter 8.

Which of the following examples of signaling pathways could be directly involved in cell differentiation? A. A steroid hormone enters the cell, interacts with its receptor and then activates transcription. B. A protein binds to a transmembrane receptor and sends a signal to the cell to enter mitosis. C. A protein binds to a transmembrane receptor and sends a signal to the cell to undergo apoptosis. D. All of the above. E. None of the above.

Role of Differential Gene Expression

Important question that scientists had to answer: o Do cells express different genes because they contain different genes or o Do cells all contain the same genes but express different subsets??? Differential gene expression: expression of different genes in different cell types o key to cell differentiation during development

Animal Cloning
Mammary gland cells from adult sheep were fused with enucleated eggs o resulting embryos were implanted into surrogate mothers o genetically identical clone of parent sheep was born (Dolly) Other species have now also been successfully cloned. Research on cloning has shown that generally (exception = certain immune cells), cellular differentiation does not involve changes in genetic makeup of cells, but rather results from differential gene expression.

Vol 445|22 February 2007

SPECIAL REPORT

Dolly: a decade on
Ten years ago, the birth of Dolly the sheep sparked a media frenzy and a prolonged ethical debate. Today, the arguments have switched focus to stem cells, and the research itself is beginning to change tack.

Article posted on blackboard!

cientists clone adult sheep triumph of the nuclear family. Activists cautioned that of UK raises alarm over human use, fertility doctors would perfect the technique ran the first headline announcing the and get rich making clones for the infertile, the cloning of an adult mammal ten years ago this narcissistic and the eccentric. US President Bill week. Ian Wilmut at the Roslin Institute near Clinton announced that the feat raises serious Edinburgh and his colleagues at PPL Thera- ethical questions and commanded his bioethpeutics in East Lothian reported on 27 Febru- ics advisers to report to him in 90 days on ary 1997 that they had produced a lamb named measures he should take to prevent its abuse. Dolly, born the previous July, that was the first Ten years later, the ethical debate launched mammalian clone created using the genetic by Dolly, and encouraged by science-fiction 1 material from an adult cell . stories, has changed. It has been supplanted by As soon as the story hit the front page (the one that is more complex, more rooted in realnews was broken by a British Sunday news- ity and far more relevant to the research that paper four days ahead of Natures publication), scientists want to do. a public and media maelstrom ensued. What didnt happen was the birth of a The first press calls came from New Zea- [cloned] child or a widespread public demand land, recalls Sue Charles of Northbank Com- for the use of cloning for reproduction, says munications in London, who was handling Alta Charo, a professor of law and bioethics publicity for Roslin and PPL at the time. They at the University of Wisconsin, Madison, who worked their way through Ausserved as a bioethics adviser to tralia, Asia and Europe. Later This is one of those Clinton. What did happen was in the day, calls began coming areas where just a complete shift in the ethical reproductive in from the United States, with trying to rein in nutty discussion from research uses. interest ranging from the sciuses of cloning to ence involved to politics and behaviour became a And a merging of the cloning religion. We even had a US full-time job. debate into the debate around chat show that wanted Dolly embryonic stem-cell research on, says Charles. They offered to fly her over. to the disadvantage of both fields because She remembers that her team, together with of the attendant confusion. Wilmut and his colleagues, took around 2,000 Back in early 1997, none of Wilmut and his calls from journalists in two weeks. colleagues, the referees who reviewed their But Dolly wasnt popular with everyone. paper, or the Nature editors who oversaw it, Pundits warned of a future in which armies of anticipated the huge public reaction to the human clones would be created by the evil and cloning of Dolly. Scientists in the field saw her egotistical. Conservatives predicted the demise birth as an incremental advance in large

Pic cap in heere

Celebrity sheep: Dolly, the first clone of an adult mammal, became an unlikely media star.

part because one year earlier, Nature had published a paper from Wilmuts group reporting the cloning of two lambs, Morag and Megan, using nuclei from embryonic cells2. I always maintained that Dolly was expected and Morag and Megan were truly surprising, says Davor Solter, director of the Max Planck Institute for Immunobiology in Freiburg, Germany. Solter wrote a News & Views article in Nature about the paper on Morag and Megan, suggesting that it was time to start thinking about the implications and uses of cloning mammals from adult cells3. Philip Campbell, Natures editor-in-chief, also recalls that the media storm over Dolly took him by surprise. Staff and referees were aware that this was the paper that in principle demonstrated how to clone mammals, including humans, he says. But neither they nor I

CLONING TIMELINE

1952
Robert Briggs and Thomas King in Philadelphia, Pennsylvania, describe how they cloned frogs (Rana pipiens) by replacing the nuclei of eggs with cells from tadpoles and adult intestinal epithelium. A similar experiment was first proposed by Hans Spemann at the University of Freiberg, Germany, in 1938.

1984
Chinese researchers clone a fish the crucian carp (Carassius carassius) from cultured kidney cells.

1996
Researchers at the Roslin Institute in Scotland clone two lambs Megan and Morag from embryonic cells. This was a crucial step towards cloning an animal from an adult cell, and is seen by some scientists as a bigger breakthrough than Dolly herself.

1997
Roslin researchers announce the birth of Dolly the sheep, the first mammal to be cloned from an adult cell, igniting public debate about the prospects for cloning humans.

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D. M. DENNIS/PHOTOLIBRARY; FLPA; ROSLIN INST.

Will the cloned sheep (Dolly) have a white or black face? A. White B. Black

How Does Differential Gene Expression Occur?

A gene can be regulated at multiple levels: o transcription o RNA processing o translation o post-translation Transcription is the fundamental level of control in differential gene expression during development. In eukaryotes, transcription is controlled primarily by the presence of proteins called regulatory transcription factors.

Developmental Biology is Interdisciplinary

Recall from Chapter 18.

Cell-Cell Signals Trigger Differential Gene Expression

Fate of a cell depends on: o timing (current stage of organisms development) o spatial location (physical location in organisms body) determined in early development by 3 major body axes: Anterior-posterior, Ventral-dorsal, Left-right Cell-cell signals tell cells where they are in time and space o signals activate transcription factors that turn specific genes on or offresulting in differentiation

Master Regulators Setup the Major Body Axes

Pattern formation: series of events that determine spatial organization of embryo Certain early signals act as master regulators o activate network of genes that send signals with more specific information about spatial location of cells sets up major body axes of the embryo

Pattern formation is progressive o as development proceeds, series of signals arrive that activate genes specifying finer control over fate of cell

Pattern Formation Mutants

1970s (Christiane Nsslein-Volhard and Eric Wieschaus) o Genetic approach to studying development in fruit fly (Drosophila melanogaster) produced mutant embryos identified more than 100 genes that play fundamental roles in pattern formation

Pattern Formation Mutant: the bicoid gene

Nsslein-Volhard and Wieschaus found dramatic mutation: o structures on anterior end were replaced with posterior structures named mutant gene bicoid, for two-tailed suspected bicoid gene product provides positional information hypothesized that bicoid coded for protein signal that tells cells where theyre located along anteriorposterior body axis

Where is the bicoid product found?

Determined location of bicoid mRNA in the egg o bicoid mRNA was highly localized in anterior of egg Bicoid protein is made from mRNA in anterior end and diffuses away from that end of the embryo produces steep concentration gradient from anterior to posterior end

How Does Bicoid Work?

Bicoid protein - regulatory transcription factor o acts as master regulator within Drosophila embryos o turns on genes responsible for forming anterior structures o absence contributes to formation of posterior structures Concentration gradient formed by Bicoid protein o provides cells with information about their position along anterior-posterior axis

Bicoid Pattern Formation Movie

Segmentation Genes
Segment: repeated regions along an animals body that define distinct animal body regions containing distinct structures o segmentation genes organize cells and tissues into distinct segments code for regulatory transcription factors

What do Segmentation Genes Do?

Three general classes of segmentation genes identified in Drosophila: o Gap genes: define general position of segments in anterior, middle, or posterior of body o Pair-rule genes: demarcate boundaries of individual segments o Segment polarity genes: delineate boundaries within individual segments Segmentation gene sets are expressed in sequence and in increasingly restricted regions o 1) Gap genes, 2) Pair-rule genes, 3) Segment polarity genes

Homeotic Genes
After segmentation genes establish identity of each segment along anterior-posterior axis, development continues with activation of homeotic genes o Segmentation gene products establish the boundaries of each segment o Homeotic gene products identify each segments structural role Trigger development of structures that are appropriate to each type of segment

Hox Genes
Eight homeotic genes in Drosophila = Hox genes o expressed in a distinctive pattern along the anteriorposterior axis, after segments are established o code for regulatory transcription factors that trigger the production of segment-specific structures

Some Drosophila mutants have segments that have been transformed into another segment, with its associated structures o occurs when cells get incorrect information about where they are in the body

Regulatory Genes Form Cascades

Interactions among bicoid and segmentation genes form a regulatory cascade Master regulators trigger production of other regulatory signals and transcription factors.which trigger production of another set of signals and regulatory proteinsand so on. The bicoid gene, gap genes, pair-rule genes, segment polarity genes, and homeotic genes each define a level in the cascade!!!

Regulatory gene cascades lead to ___________. A. Transcription of genes coding for certain enhancers. B. Transcription of genes coding for certain regulatory transcription factors. C. Binding of proteins to certain promoters. D. All of the above. E. None of the above.

Overall Function of Regulatory Genes

Regulatory genes act in a sequence, triggering gene cascades that provide progressively detailed information about where cells are located in time and space o Cells receive unique positional information because identity and concentration of transcription factors vary along the three major body axes o Each level in a regulatory cascade provides a more specific level of information about where a cell is o As regulatory cascades proceed, a cell's fate becomes more and more finely determined!

Concepts to Remember
During development, cells divide, move in a directed manner, become specialized (differentiated), and interact with other cells. Cells become specialized because they express different genes, not because they contain different genes. (Review Chapter 18) Cells interact continuously during development by means of cell-cell signals. (Review Chapter 8) When development begins, early cell-cell signals trigger a cascade of effects that cause increasing specialization as development proceeds.