Adverse effects of serotonin depletion in developing zebrafish.

In this study, p-chlorophenylalanine (pCPA), an inhibitor of tryptophan hydroxylase (the rate limiting enzyme of serotonin synthesis), was used to reduce serotonin (5HT) levels during early development in zebrafish embryos. One day old dechorionated embryos were treated with 25 µM pCPA for 24h and subsequently rescued. Immunohistological studies using a 5HT antibody confirmed that 5HT neurons in the brain and spinal cord were depleted of transmitter by 2 days post fertilization (dpf). Twenty four hours after pCPA exposure embryos were unable to burst swim and were nearly paralyzed. Movement began to improve at 4 dpf, and by 7 dpf, larvae exhibited swimming activity. Rescued larvae continued to grow in rostrocaudal length over 5 days post-rescue, but their length was always 16-21% below controls. Surprisingly, both groups displayed the same number of myotomes. To examine whether hypertonicity of myotomes in treated embryos played a role in their shorter rostrocaudal lengths, 1 dpf embryos were exposed to a combination of 25 µM pCPA and 0.6 mM of the sodium channel blocker ethyl 3-aminobenzoate methanesulfonate (MS-222). After a 24 hour exposure, the embryos exhibited the same rostrocaudal length as control embryos suggesting that myotome hypertonicity plays a major role in the decreased axial length of the treated larvae. In addition, pCPA treated 2 dpf embryos exhibited abnormal notochordal morphology that persisted throughout recovery. Reverse transcriptase polymerase chain reaction (RT-PCR) was performed to determine the relative levels of the serotonin 1A receptor (5HT(1A)) transcript and the serotonin transporter (SERT) transcript in the brain and spinal cord of control and treated embryos. Transcripts were present in both brain and spinal cord as early as 1 dpf and reached maximal concentrations by 3 dpf. Embryos treated with pCPA demonstrated a decrease in the concentration of 5HT(1A) transcript in both brain and spinal cord. While SERT transcript levels remained unaffected in brain, they were decreased in spinal cord. Five days subsequent to pCPA rescue, 5HT(1A) transcript concentrations remained decreased in brain while SERT transcript levels were elevated in both regions. These findings suggest that reduction of 5HT during early zebrafish development may have an adverse effect on body length, notochordal morphology, locomotor behavior, and serotonin message-related expression.

Dose-response effects of chronic lithium regimens on spatial memory in the black molly fish.

Lithium is widely used in the management of bipolar disorder, yet memory impairment is a serious side effect. To assess the effects of lithium on spatial working and reference memories, we have employed a plus maze utilizing spontaneous alternation (SA) and place-learning paradigms in two experiments with the black molly fish. Four treatment groups were gavaged with 20 microl of a 10, 100, or 1000 mM lithium chloride (LiCl) solution or ddH(2)O vehicle every 12 h for 22 to 24 days. On Day 15, subjects began an 8-day SA task or a 10-day place-learning task. Results indicate that there is a significant difference in SA performance among the treatment groups for Days 1, 2, and 3. Results of the place-learning task indicate that the 1 M dose group needed significantly more trials to reach criterion and made significantly fewer correct first choices than the other dose groups. Capillary ion analysis determinations of plasma and brain lithium levels illustrate linear dose-response relationships to doses administered. Regression analyses indicate that there is a relationship between SA performance and plasma/brain lithium levels during the initial part of testing. Collectively, the results indicate that chronic lithium administration impairs spatial working and reference memories.

Comparative study of dentition among species of Poecilia (Pisces).

Many studies in the genus Poecilia have focused on reproductive and genetic characteristics of Poecilia formosa, the Amazon molly, and its sympatric species P. latipinna and P. mexicana. The research literature of Poecilia dentition has been limited to general tooth morphology. Essentially absent are comparative analyses of dentition patterns and total numbers of teeth. The current study uses dentition analysis as a method to compare species in the genus Poecilia and to address some taxonomic issues related to these fish. The study focused on fish from the areas of southern Texas and northeastern Mexico. Through the use of scanning electron microscopy, the lower jaws of Poecilia spp were examined to determine total numbers of outer and inner teeth. In addition, the differences in distribution patterns of the inner teeth were recorded and compared. Statistical analyses were performed to determine which comparisons were significant. This study reveals several observations: 1) variations in the numbers of outer and inner teeth exist in some of these fish with respect to site of collection; 2) differences in total teeth numbers and dentition patterns were found both interspecifically and intraspecifically; and 3) in addition, dentition analysis provided evidence regarding the origin of P. formosa. This study supports the current notion that P. latipinna, the proported paternal component, and P. mexicana limantouri, the purported maternal component, are the progenitor species of P. formosa. Two unresolved taxonomic questions were addressed through dentition analysis. First, the present study supports the exclusion of the triploid associate of P. formosa as a separate species from P. formosa. Second, this study shows a significant difference in the number of inner teeth and in dentition patterns between P. mexicana limantouri and P. mexicana mexicana. Such differences, in addition to previously known distinguishing characteristics, should prompt careful consideration of whether or not these taxa deserve specific status or retention of their current subspecific status. J. Morphol. 239:271-282, 1999. © 1999 Wiley-Liss, Inc.