An Introduction to Genetic Statistics.

There is increasing interest in determining the extent to which multiple characters related to drug sensitivity are influenced by common genes. The principal method for testing for the existence of such genetic correlations has been examination of pairs of mouse or rat lines selectively bred for sensitivity or resistance to a single behavioral effect of a drug. When a pair of selected lines is found to differ significantly on some trait other than the one on which they were selected, it is commonly concluded that significant genetic correlation between the traits exists, implying the action of a common set of genes on the two responses. In addition, results from comparisons of lines of animals selected for trait X and tested for trait Y may be compared with results from lines selected for trait Y and tested for trait X. As the number of correlated responses in selected lines increases, it becomes more important to adhere to sensible, consensual guidelines for interpreting such line differences. The principles underlying phenotypic and genotypic correlational analyses with selected lines are discussed. A scheme is presented to allow standardization across laboratories of inferences about the relative strength of genetic association from experiments with selected lines. Statistical and practical experimental issues are addressed. Estimates of genetic correlations may also be derived from the correlation of mean trait values across a panel of inbred strains. Existing data have sometimes found estimates of genetic correlations made with one approach to be inconsistent with those estimated in other ways. Possible reasons for this are discussed. Finally, the relationship between phenotypic correlations and genetic correlations is discussed. Phenotypic and genetic correlations for a pair of traits may differ widely, and may even be opposite in sign. Both are characteristic of the population from which they are sampled. Phenotypic correlations estimated within selected lines may change over time, as the additive genetic variance in the selected trait is exhausted. A specific example of this phenomenon is given.

Estimation of Genetic Correlation: Interpretation of Experiments Using Selectively Bred and Inbred Animals

Fetal rat spinal cord tissue was obtained on gestational day 14 (E14) and transplanted into 2-4-mm-long intraspinal cavities produced by partial spinal cord lesions in adult and neonatal rats. At regular post-transplantation intervals, light and electron microscopy, autoradiographic demonstration of tritiated thymidine labelling, and immunocytochemical localization of glial fibrillary acidic protein (GFAP) were used to identify surviving donor tissues and to study their differentiation and extent of fusion with recipient spinal cords. In some experiments, wheat germ agglutinin-conjugated horseradish peroxidase (WGA-HRP) was also employed to examine whether neurons within the grafts projected axons into the host spinal cord and vice versa. Lastly, immunocytochemistry was used to determine whether any supraspinal serotoninergic (5-HT) axons from the host extended into the transplants. Over 80% of the grafts survived in lesions of both the neonatal and adult rat spinal cord for periods of 1-16 months (duration of experiment), and considerable maturation of donor tissue was evidenced, which even included the appearance of some topographical features of the normal spinal cord. Many of the transplants extended the entire length of the lesion, and were often closely apposed to the injured surfaces of the recipient spinal cords without an intervening dense glial scar. At post-transplantation intervals of 2-4 months, injection of WGA-HRP into the host spinal cord (5 mm from the transplant in adult animals or as much as 20 mm in neonatal recipients) demonstrated retrogradely labelled neurons and anterogradely labelled axons in the grafts. Likewise, injecting WGA-HRP into transplants in adult recipients resulted in labelling of neurons in adjacent segments of the host spinal cord; some labelled axons, derived from donor neurons, were also present in neighboring spinal gray matter. Finally, immunocytochemistry revealed 5-HT-like immunoreactive fibers in transplants that had been prelabelled with tritiated thymidine. These observations demonstrate the potential of embryonic spinal cord transplants to replace damaged intraspinal neuronal populations and to restore some degree of anatomical continuity between the isolated rostral and caudal stumps of the injured mammalian spinal cord.

Intraspinal transplantation of embryonic spinal cord tissue in neonatal and adult rats.

Towards an improved serum-free, chemically defined medium for long-term culturing of cerebral cortex tissue

Publisher Summary This chapter focuses on an in oculo technique with emphasis on technical details and step-by-step procedures and presents an analysis of various types of grafts. The two most obvious advantages of the in oculo approach are that it provides a chamber in which grafts can exist without being squeezed by or exerting pressure on host tissues and the transparency of the cornea through which grafts can be noninvasively observed. The grafted tissue that acts as a receptor of nerves from the host iris may itself be nervous tissue. The chapter presents a demonstration of partial phenotypic transformation of adult chromaffin cells to neuron-like cells as fiber outgrowth from chromaffin cells grafted to sympathetically denervated irises. Intraocular grafting, when properly performed, is probably among the least harmful types of experimental surgery that can be performed on a rat.

Intraocular Transplantation in Rodents: A Detailed Account of the Procedure and Examples of its Use in Neurobiology with Special Reference to Brain Tissue Grafting

In the present study, we compared phenotypic differences in behavioral and neurophysiological responses to acute ethanol administration among eight inbred strains of mice. Genetic variation for behavioral sedation, as measured by loss of the righting reflex (sleep time) after a hypnotic dose of ethanol, was shown to be present among the inbred strain population. In addition, a large genetic component of variation in the depressant action of ethanol on the spontaneous discharge of cerebellar Purkinje neurons was found. Results from an analysis of covariance of the behavioral and electophysiological trophysiological phenotypes, measured on each mouse among the inbred strains, provided strong evidence for a high genetic correlation between sleep time and inhibition of cerebellar Purkinje neuron discharge in response to acute ethanol administration. Taken together with our previously reported data on ethanol-induced electrophysiological changes in selectively bred lines, the results described here strongly support the hypothesis that the cerebellar Purkinje neuron is one important locus for the acute soporific effects of alcohol.

Evidence for genetic correlation of hypnotic effects and cerebellar Purkinje neuron depression in response to ethanol in mice

Trophic effects of brain areas on the developing cerebral cortex: II. Electrophysiology of intraocular grafts

Survival and growth of neurons with enkephalin-like immunoreactivity in fetal brain areas grafted to the anterior chamber of the eye.

The anatomical and electrophysiological basis of enkephalin-catecholamine interactions was studied in the central nervous system of the Sprague-Dawley rat. Sections of fetal rat brain were treated alternatively with antibodies raised against enkephalin or tyrosine hydroxylase (EC 1.14.16.2). Enkephalinergic fibers were found to innervate almost all brain areas known to contain catecholamine cell bodies or terminal fields. The possible electrophysiological correlates of such an overlap in enkephalin and catecholamine terminal fields was investigated in frontal neocortex. Parenteral administration of antipsychotic agents known to block catecholamine receptors, such as spiroperidol, alpha-flupenthixol, and (+)-butaclamol, reversibly antagonized enkephalin-induced depressions of neuronal activity. The biochemically inactive isomers of these antipsychotics, beta-flupenthixol and (-)-butaclamol, manifested no such activity. Unilateral lesions of the catecholaminergic projections to frontal cortex produced by interstitial injection of 6-hydroxydopamine resulted in an ipsilateral decrease in enkephalin efficacy and in an elimination of the antagonisms of enkephalin depressions by antipsychotic agents. It is hypothesized that catecholamines may act as modulators to augment responsiveness to peptides such as enkephalins.

M.R. Palmer

Papers

Evidence for genetic correlation of hypnotic effects and cerebellar Purkinje neuron depression in response to ethanol in mice

Trophic effects of brain areas on the developing cerebral cortex: II. Electrophysiology of intraocular grafts

Survival and growth of neurons with enkephalin-like immunoreactivity in fetal brain areas grafted to the anterior chamber of the eye.

Modulatory interactions between enkephalin and catecholamines: anatomical and physiological substrates.

Postnatal lead exposure alters spontaneous cerebellar Purkinje neuron discharge