Jose R. Eguibar

Lordosis facilitation in estrogen primed rats by intrabrain injection of pregnanes

Progesterone (P) and nine of its natural metabolites were bilaterally injected (5 micrograms in 0.5 microliter oil) into either the ventromedial hypothalamus (VMH) or the medial preoptic area (MPOA) of estrogen primed rats to assess their relative potencies for stimulating lordosis. P, 5 alpha-pregnanedione and 5 beta, 3 beta-pregnanolone elicited lordosis when injected at either VMH or MPOA. By contrast, 5 alpha, 3 beta-pregnanolone as well as 20 alpha-OH and 20 beta-OH-pregnenone were much more effective in stimulating lordosis when implanted in the MPOA. Finally, 5 beta-pregnanedione and 5 beta,3 alpha-pregnanolone did not stimulate lordosis at neither VMH nor MPOA. The observation that lordosis was induced in estrogen primed rats both by pregnanes that bind to the P receptor (i.e., P; 5 alpha-pregnanedione; 20 alpha- and 20 beta-OH-pregnenone) and by pregnanes that do not (i.e., 5 alpha, 3 beta-; 5 beta,3 beta- and 5 alpha,3 alpha-pregnanolone) indicates that diverse cellular mechanisms are involved in the facilitation of lordosis by pregnanes.

Neurophamacology of Yawning

Yawning is a common behavioral event that is observed in humans, as well as other mammals, birds and reptiles. In humans, yawning often occurs just before bed and upon waking up, and is also associated with tedious or boring situations. Although the physiologic roles of yawning have yet to be fully elucidated, the past 50 years of research has led to a much greater understanding of the neuropharmacologic regulation of yawning. While many of the early studies concluded that yawning was primarily driven by changes in cholinergic neurotransmission, we now know that numerous neurotransmitters and neurohormones are involved in the mediation of yawning, including acetylcholine, dopamine, glutamate, serotonin, oxytocin, GABA, opioids, adrenergics, nitric oxide, as well as the proopiomelanocortin-derived peptides ACTH and alpha-MSH. Furthermore, antagonist interaction studies have clearly defined at least 3 distinct neural pathways involved in the induction of yawning, as well as the hierarchical order through which these different neurotransmitter systems interact to regulate yawning. The following sections will discuss the state of knowledge for each of the major neurotransmitters and neurohormones involved in the regulation of yawning, their interactions with one another, and their place in the hierarchical organization of yawning.

Behavioral differences between selectively bred rats: D1 versus D2 receptors in yawning and grooming.

We used SKF 38393 and quinpirole for determining whether activation of D(1) and D(2) receptors, respectively, is involved in behaviors of rats selectively bred for high or low rates of yawning. After injection of SKF 38393, yawning diminished more markedly in high-yawning (HY) than in low-yawning (LY) rats, whereas this drug increased the number and duration of grooming episodes similarly in both strains. After injection of quinpirole, yawning increased more markedly in HY than in LY rats, whereas this drug decreased the number and duration of grooming episodes similarly in both rat strains. After coadministration of SKF 38393 and quinpirole, yawning increased similarly in both rat strains, whereas the combination of drugs failed to reliably affect grooming behavior. We interpret our findings as indicating that D(2) receptors are more important than D(1) receptors for differences in yawning behavior between HY and LY rats.

Progressive Deterioration of Central Components of Auditory Brainstem Responses during Postnatal Development of the Myelin Mutant taiep Rat

Auditory brainstem responses (ABRs) were evaluated during the postnatal development (P10–P180) of taiep rats, neurological mutants characterized by early abnormal myelin development and subsequent demyelination of the CNS. The disorder is produced by an autosomal recessive mutation trait that affects the oligodendrocytes but not the Schwann cells. After onset of ABRs (P12–P14), taiep rats and their nonaffected heterozygous littermates that served as controls showed a similar pattern of maturation for wave I. The central waves (In–IV) showed significantly longer latencies in the mutants. By P60–P180, the later waves (III and IV) were frequently difficult to discern. From the onset of ABRs, the interpeak latency I–IV, corresponding to the central conduction time (CCT) of the auditory pathway, showed in taiep rats significantly longer values than controls. After an initial reduction, proportional to that of control rats, the CCT value increased progressively during the second month of the mutants’ lives. The electrophysiological results of the present study strongly support the hypothesis that mutation in the taiep rat impairs neuromaturation of the central auditory pathway in the brainstem by affecting the myelination process in the CNS.

Regional and temporal progression of reactive astrocytosis in the brain of the myelin mutant taiep rat.

Reactive astrocytosis in taiep rats was shown by glial fibrillary acidic protein (GFAP) immunoreactivity measured by means of enzyme-linked immunosorbent assay and indirect immunofluorescence. Increased GFAP immunoreactivity was first observed in the brainstem of 15-day-old taiep rats and was widespread throughout all brain regions at 6 months of age. Characteristically, astrocytes were hypertrophic and displayed strong GFAP fluorescence. The pattern of these reactive cells may correlate with the process of dysmyelination in the taiep rat.

Copulation and ejaculation in male rats under sexual satiety and the Coolidge effect

Sexually satiated males cease copulating after several ejaculations with the same female; and the presence of an unknown receptive female renews copulation including ejaculation, a process named the Coolidge effect. It is believed that the Coolidge effect has the aim to impregnate another female, although it is known that the sperm count gradually decreases after consecutive ejaculations. The main goal was to investigate if sexually satiated males during the Coolidge effect can reestablish seminal expulsion associated to the ejaculation behavior and/or penile erection associated to the intromission behavior. The results show that during the Coolidge effect, most of the sexually satiated males showed the motor ejaculatory behavior, however, no sperm in the uterine horns or seminal plug in the vagina were detected. Such lack of sperm was not related with the number of ejaculations required to achieve sexual satiety nor with the number of intromissions needed for ejaculating (experiment 1: 2.4.1.). After the behavioral ejaculation, during the Coolidge effect, there was a 44% decrease in sperm count in the epididymal caudae (experiment 1: 2.4.2.). Males that mated for 8 behavioral ejaculations (close to sexual satiety) deposited tiny seminal plugs but no sperm in the female reproductive tract (experiment 1: 2.4.3.). Interestingly, sexually satiated and non-satiated-animals displayed similar number of intromissions and spent a similar time in dislodging the seminal plug from the vagina deposited by other males (experiment 2). These results suggest that sexually satiated males during the Coolidge effect have the capacity for penile erection and vaginal insertion, because they are able to dislodge seminal plugs; but are unable to expel seminal fluid, because neither form seminal plugs nor deposit sperm in the female genital tract.

Smell facilitates auditory contagious yawning in stranger rats

Most vertebrates yawn in situations ranging from relaxation to tension, but only humans and other primate species that show mental state attribution skills have been convincingly shown to display yawn contagion. Whether complex forms of empathy are necessary for yawn contagion to occur is still unclear. As empathy is a phylogenetically continuous trait, simple forms of empathy, such as emotional contagion, might be sufficient for non-primate species to show contagious yawning. In this study, we exposed pairs of male rats, which were selected for high yawning, with each other through a perforated wall and found that olfactory cues stimulated yawning, whereas visual cues inhibited it. Unexpectedly, cage-mate rats failed to show yawn contagion, although they did show correlated emotional reactivity. In contrast, stranger rats showed auditory contagious yawning and greater rates of smell-facilitated auditory contagious yawning, although they did not show correlated emotional reactivity. Strikingly, they did not show contagious yawning to rats from a low-yawning strain. These findings indicate that contagious yawning may be a widespread trait amongst vertebrates and that mechanisms other than empathy may be involved. We suggest that a communicatory function of yawning may be the mechanism responsible for yawn contagion in rats, as contagiousness was strain-specific and increased with olfactory cues, which are involved in mutual recognition.

Leptin facilitates lordosis behavior through GnRH-1 and progestin receptors in estrogen-primed rats

Dose response curves for leptin facilitation of estrous behavior (lordosis and proceptivity) were made by infusing the peptide into the lateral ventricle (icv) of ovariectomized (ovx), ad libitum-fed rats injected 40h previously with 5μg of estradiol benzoate. Leptin doses of 1 and 3μg produced significant lordosis quotient at 60min post-injection, with maximal lordosis being displayed at 120min. Yet the intensity of lordosis was weak, and a high incidence of rejection behaviors was found. Moreover, leptin did not induce significant proceptive behaviors at any dose. The leptin doses of 1 and 3μg were selected for determining whether antide, a GnRH-1 receptor antagonist, or the progestin receptor antagonist RU486 could modify the lordosis response to leptin. Icv injection of either antide or RU486 1h before leptin significantly depressed leptin facilitation of lordosis. The results suggest that leptin stimulates lordosis by releasing GnRH, which in turn activates GnRH-1 and progestin receptors. The physiological role of leptin in the control of estrous behavior remains to be determined.

The nitric oxide pathway participates in lordosis behavior induced by central administration of leptin

Intracerebroventricular (icv) administration of leptin facilitates lordosis behavior in ad libitum-fed, estrogen-primed rats. The cellular mechanism involved in this response is unknown. The present study tested the hypothesis that the nitric oxide-guanylyl cyclase, cGMP-dependent protein kinase (PKG) pathway is involved in the facilitation of lordosis behavior induced by the central administration of leptin. We tested the importance of the nitric oxide/cGMP pathway for lordosis stimulation by either icv infusion of a nitric oxide synthase inhibitor (L-NAME) or a nitric oxide-dependent, soluble guanylyl cyclase inhibitor (ODQ) 30 min before leptin administration (1 μg). This dose of leptin reliably induced lordosis behavior in ovariectomized estradiol benzoate treated rats. The lordosis induced by leptin at 1 and 2h after infusion was significantly reduced by the previous injection of either L-NAME or by ODQ. Intracerebroventricular infusion of the PKG inhibitor (KT5823) 30 min before leptin infusion, also significantly inhibited the lordosis behavior induced by leptin at 1 and 2h after hormone administration. These data support the hypothesis that the nitric oxide/cGMP/PKG pathway is involved in the facilitation of lordosis by leptin in estrogen-primed female rats.

Developmental impairment of compound action potential in the optic nerve of myelin mutant taiep rats.

The taiep rat is a myelin mutant with an initial hypomyelination, followed by a progressive demyelination of the CNS. The neurological correlates start with tremor, followed by ataxia, immobility episodes, epilepsy and paralysis. The optic nerve, an easily-isolable central tract fully myelinated by oligodendrocytes, is a suitable preparation to evaluate the developmental impairment of central myelin. We examined the ontogenic development of optic nerve compound action potentials (CAP) throughout the first 6 months of life of control and taiep rats. Control optic nerves (ON) develop CAPs characterized by three waves. Along the first month, the CAPs of taiep rats showed a delayed maturation, with lower amplitudes and longer latencies than controls; at P30, the conduction velocity has only a third of the normal value. Later, as demyelination proceeds, the conduction velocity of taiep ONs begins to decrease and CAPs undergo a gradual temporal dispersion. CAPs of control and taiep showed differences in their pharmacological sensitivity to TEA and 4-AP, two voltage dependent K+ channel-blockers. As compared with TEA, 4-AP induced a significant increase of the amplitudes and a remarkable broadening of CAPs. After P20, unlike controls, the greater sensitivity to 4-AP exhibited by taiep ONs correlates with the detachment and retraction of paranodal loops suggesting that potassium conductances could regulate the excitability as demyelination of CNS axons progresses. It is concluded that the taiep rat, a long-lived mutant, provides a useful model to study the consequences of partial demyelination and the mechanisms by which glial cells regulate the molecular organization and excitability of axonal membranes during development and disease.