Histological aspects of in situ hybridization

Summary This study examined the detection of cellular poly(A) sequences in mouse liver sections by in situ hybridization using a ³H-labelled poly(dT) probe. Parameters examined included possible losses of target poly(A) sequences from sectioned cells, access of probe to target sequences, section thickness, hybridization conditions, autoradioigraphic efficiency, specific activity of probes and specificity of reaction. An improved protocol was devised that resulted in good preservation of histological detail in sectioned tissue blocks, and a calculated hybridization efficiency of 50%–100%. With the use of probes of defined sequence, the protocol should allow detection of unique mRNA sequences within single cells with an estimated sensitivity of 6–12 unique mRNA molecules per sectioned cell.     

Learning Influences Courtship Preferences of Male Threespine Sticklebacks (Gasterosteus aculeatus)

Male threespine sticklebacks (Gasterosteus aculeatus) prefer to court the more gravid of a pair of dummy females (Rowland 1982, 1989). In this study, males were trained to modify their courtship preferences. Males were given two pre-punishment preference trials during which two dummy females—one approximating a normally gravid female and the other, a supergravid female, possessing an abnormally large abdomen — were simultaneously presented to males. After pre-punishment trials, males were subjected to one of two punishment procedures: punishment with weak electric shock (Experiment I) or punishment by the removal of females to simulate female retreat (Experiment II). During punishment trials, males were punished each time they courted the supergravid dummy. Control males were subjected to identical trials, but were not punished for courting the supergravid dummy. Post-punishment preference trials were given to all males (Experiments I and II). The results of Experiments I and II were similar. Punished and control males did not differ in their courtship preferences in pre-punishment trials; they preferred to court the supergravid dummy. Punished males, however, spent a significantly lower proportion of time visiting and directed a smaller proportion of zigzags (a courtship display) toward the supergravid dummy than did control males during punishment training (Experiment I) and post-punishment preference trials (Experiments I and II). These results suggest that courtship preferences of male threespine sticklebacks can be modified through learning. Such a mechanism could be important for adaptive mate choice under natural conditions.     

Timing of parturition in the rat: an analysis of successive births

To determine whether consecutive parturitions within individual rats are likely to occur around the same time of the 24h day, female rats mated prior to ovulation and maintained on a LD 12:12 photoperiod, were observed over either two or three parturitions. A subgroup of females was placed on constant light during the second gestation in order to determine the role of photoperiod in the temporal synchronization of consecutive births. The major findings were 1. individual females tended to give birth around the same time of the day on consecutive births so long as the photoperiod remained constant; 2. the small percentage of births occurring during darkness, the less-prevalent time of the photoperiod for birthing, were anomalous births for individual females; and 3. litter weight was a significant factor in predicting time of birth for females, but this effect interacted with photoperiod. These data elucidate the importance of knowing previous birth time, litter weight, and photoperiod on predicting the time of parturition in the rat.     

Synthesis and characterisation of D-amino acid-based oligopeptides for use as probes of the influence of molecular structure on the paracellular route of gastrointestinal drug uptake

Summary A series of peptides based on D-amino-acids, and with an N-terminal D-phenylalanine residue, was synthesised by solution methods using t-butoxycarbonyl amino protection. These peptides were designed to resist metabolism in the gastrointestinal tract and to serve as probes of the effects of molecular shape and charge on the paracellular route of drug uptake in the gut. The peptides were characterised by NMR spectroscopy, FAB mass spectrometry, optical rotation and purified by HPLC.Abbreviations HBTU o-Benzotriazolyl-tetramethyluronium hexafluorophosphate - OSu N-hydroxysuccinimide ester