Characterization of Cre recombinase mouse lines enabling cell type-specific targeting of postnatal intervertebral discs

Cre/loxP technology is an important tool for studying cell type-specific gene functions. Cre recombinase mouse lines, including Agc1-CreER^T2, Col2a1-Cre; Col2a1-CreER^T2, Shh-Cre, Shh-CreER^T2, and Osx-Cre, have been proven to be valuable tools to elucidate the biology of long bones, yet the information for their activity in postnatal intervertebral disc (IVD) tissues was very limited. In this study, we used R26-mTmG fluorescent reporter to systematically analyze cell specificity and targeting efficiency of these six mouse lines in IVD tissues at postnatal growing and adult stages. We found that Agc1-CreER^T2 is effective to direct recombination in all components of IVDs, including annulus fibrosus (AF), nucleus pulposus (NP), and cartilaginous endplate (CEP), upon tamoxifen induction at either 2 weeks or 2 months of ages. Moreover, Col2a1-Cre targets most of the cells in IVDs, except for some cells in the outer AF (OAF) and NP. In contrast, the activity of Col2a1-CreER^T2 is mainly limited to the IAF of IVD tissues at either stage of tamoxifen injection. Similarly, Shh-Cre directs recombination specifically in all NP cells, whereas Shh-CreER^T2 is active only in a few NP cells when tamoxifen is administered at either stage. Finally, Osx-Cre targets cells in the CEP, but not in the NP or AF of IVDs tissues at these two stages. Thus, our data demonstrated that all these Cre lines can direct recombination in IVD tissues at postnatal stages with different cell type specificity and/or targeting efficiency, and can, therefore, serve as valuable tools to dissect cell type-specific gene functions in IVD development and homeostasis.     

Effect of new founders on retention of gene diversity in captive populations: A formalization of the nucleus population concept

A nucleus population is a small captive population genetically supported by periodic importation of wild caught animals. Periodic importation will allow nucleus populations to maintain the same amount of gene diversity as larger captive populations that do not import wild caught animals. The function of nucleus populations as envisioned by the IUCN/SSC Captive Breeding Specialist Group (CBSG) is to make additional captive space available for endangered taxa not currently maintained in captivity. In this article, mathematical models are developed to assess the effectiveness of the nucleus population concept in reducing the population sizes necessary to maintain appreciable amounts of gene diversity in captive populations. It is shown that the Nucleus I population concept, as defined and promoted by the CBSG, requires an importation rate 10–20 times greater than they have indicated. Whereas nucleus populations are not appropriate for maintenance of significant amounts of gene diversity in long-term breeding programs, small populations can be valuable for research, education, and reintroduction projects with short-term goals. Decisions have to be made on which of the many endangered taxa will be maintained and for what purposes, if captive breeding is to be an effective component of species conservation. © 1993 Wiley-Liss, Inc.     

The facets of olfactory learning

Volatile chemicals in the environment provide ethologically important information to many animals. However, how animals learn to use this information is only beginning to be understood. This review highlights recent experimental advances elucidating olfactory learning in rodents, ranging from adaptations to the environment to task-dependent refinement and multisensory associations. The broad range of phenomena, mechanisms, and brain areas involved demonstrate the complex and multifaceted nature of olfactory learning.     

The lateral cervical nucleus in the cat

Summary Profiles of 14 neurons all sectioned through the nucleolar plane and 87 isolated dendritic profiles have been analyzed with respect to the surface area covered by boutons and astroglial processes. This analysis has revealed two different types of neurons within the lateral cervical nucleus (LCN) of the cat. The cell types also differ in other ultrastructural respects. One type, which probably consists of projection neurons, is characterized by a rather large size, a relatively small nucleus, numerous mitochondria, well developed granular and agranular endoplasmic reticulum. The cell membrane of these cells shows somatic spines and the perikaryon is covered with boutons to a mean extent of 42%. The other cell type, which probably is internuncial, is smaller, has a proportionally larger nucleus, few mitochondria and a poorly developed granular and agranular endoplasmic reticulum. These cells show no somatic spines and the perikaryal membrane is covered with boutons to an extent of about 10%. Also the bouton populations contacting the two cell types differ from one another. The proportion of internuncial neurons within the LCN has been estimated to about 8%. The internuncial neurons seem to have no preferential localization.The primary dendrites of the projection neurons have a bouton covering of about 48%. No proportional differences in covering could be revealed between different sizes of dendrites.The results are discussed in relation to what is known about the anatomical and physiological organization of the LCN, and also compared with the results obtained in other similar investigations on other parts of the central nervous system.The author is grateful to fil.kand. Göran Engholm for his help with the statistical considerations.This work was supported by grants from Anders Otto Swärds Stiftelse, Stiftelsen Lars Hiertas minne, Åhlén och Holms stiftelse, Åke Wibergs stiftelse and the Swedish Medical Research Council (Project No B70-12X-2710).     

Aqueduct Occlusion Does Not Impair Feeding Induced by Either Third or Fourth Ventricle Galanin Injection

Exogenous galanin stimulates feeding when injected into forebrain and hindbrain sites, including the third and fourth ventricles (3V and 4V), amygdala, paraventricular nucleus of the hypothalamus (PVN), and nucleus of the solitary tract (NTS). Because the PVN and NTS border the ventricular space, it is possible that feeding stimulated by injection of galanin at these sites may be caused by the transport of galanin through the ventricular system to a remote site of action. The role of ventricular transport of galanin between the 3V and 4V in galanin-induced feeding was examined in this study. Rats were implanted with two guide cannula assemblies: one dorsal to the mesencephalic aqueduct and the other in the 3V or 4V. Feeding in response to 3V or 4V galanin injection was first measured after sham-occlusion of the aqueduct. Subsequently, flow of cerebrospinal fluid between the forebrain and hindbrain ventricles was acutely interrupted by injection of a silicone grease plug into the mesencephalic aqueduct just before assessment of the feeding response to 4V or 3V galanin injection. Aqueduct occlusion did not alter the feeding induced by either 3V or 4V galanin injection, indicating that galanin terminals in both the diencephalon and hindbrain are involved in control of food intake.     

GRF-induced feeding: Evidence for protein selectivity and opiate involvement

Growth hormone-releasing factor (GRF) is a hypothalamic peptide named for its ability to induce release of growth hormone from the anterior pituitary. GRF also acts as a neurotransmitter in the suprachiasmatic nucleus/medial preoptic area (SCN/MPOA) to stimulate food intake. The purpose of this series of experiments was to explore the nature of GRF-induced feeding, with a particular emphasis on macronutrient selectivity, and to examine the role of opiate activity in the paraventricular nucleus of the hypothalamus (PVN). Chow intake stimulated by GRF microinjection (1 pmol/0.5 μl) into the SCN/MPOA was blocked by injection of methyl-naltrexone (3 μg/0.5 μl) into the PVN. In animals habituated to macronutrient diets (Teklad, WI), GRF preferentially stimulated intake of protein at 2 and 4 h postinjection, whereas it had no effect on carbohydrate intake. Further, this effect was blocked by injection of naloxone (40 nmol/0.5 μl) into the PVN. Microinjection of morphine (0, 1, 10, and 17 μg/0.5 μl) into the PVN also specifically stimulated protein intake at 2 and 4 h postinjection. These results suggest that feeding derived from GRF actions in the SCN/MPOA is macronutrient selective, and is dependent on PVN opiate activity for expression.     

Mauthner cell-evoked synaptic actions on pacemaker medullary neurons of a weakly electric fish

The present study was designed to examine the synaptic events in neurons of the pacemaker nucleus of Gymnotus carapo during the increase in rate of the electric organ discharge following activation of Mauthner cells. Pacemaker and relay cells were investigated using intracellular recordings which were performed under two different conditions: (1) with the pacemaker nucleus spontaneously discharging and (2) after its activity was abolished by anesthesia. Mauthner axon activation induced an increase in the rate of pacemaker cell discharges. This response was accompanied by an increase in the slope of the pacemaker potential (up to 110%) and a depolarization of these cells. The discharges of relay cells followed one to one those of pacemaker cells. In contrast to that observed in pacemaker cells, only brief depolarizing antidromic effects could be evoked in relay cells after Mauthner axon activation. In quiescent pacemaker cells, Mauthner cell activation induced a prolonged (up to 500 ms) depolarizing potential with an average amplitude of 1.92 ± 0.82 mV; its latency was 4.43 ± 1.14 ms. Our data indicate that, within the pacemaker nucleus, the population of pacemaker cells is the only target for Mauthner cell-evoked, short-latency excitatory synaptic actions. Accepted: 1 March 1997     

The endosperm development and the variations of structures of embryo sacs: unravelling the low fecundity of Zephyranthes candida (Amaryllidaceae)

To unravel a low fecundity in Zephyranthes candida (Lindl.) Herb., the development of the endosperm was studied using conventional paraplast section technique. The results show that the endosperm develops normally and comprises four major stages viz. syncytial, cellularization, differentiation and maturation. Both proliferation of antipodal cells and their close contact with the primary endosperm nucleus were observed, which should favor transportation of nutrients and accelerate development of embryo and endosperm. In Z. candida, at least four events of nuclear migration occurred during the course of embryogenesis and endosperm development. The 12.7% structurally and functionally abnormal ovules, along with the 22.3% collapsed and aborted ovules observed accounts for the low fecundity to some extent.     

Dorsal and Ventral Hippocampal Sharp-Wave Ripples Activate Distinct Nucleus Accumbens Networks

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