Bruce W. Newton

Ethanol-induced alterations of neurotrophin receptor expression on Purkinje cells in the neonatal rat cerebellum

Ethanol causes loss of Purkinje cells in the cerebellum during the early stages of differentiation and maturation by a presently unknown mechanism. Neuronal vulnerability in the cerebellum parallels the prominent temporal and anatomical gradients of development (i.e. early to late interlobular and posterior to anterior, respectively). Development of Purkinje cells is known to require binding of the neurotrophins, including brain-derived neurotrophic factor (BDNF) and neurotrophin-3 (NT3), to the tyrosine-kinase (Trk) receptors TrkB and TrkC, respectively. In addition, Purkinje cells are reported to experience a critical switch between BDNF dependence and NT3 dependence during the period of highest ethanol sensitivity between postnatal days (PN) 4-6. To test the hypothesis that ethanol alters neurotrophin signaling leading to Purkinje neuronal death, the immunohistochemical expression of TrkB and TrkC receptors on Purkinje cells of rat pups following a moderate dose of ethanol was determined at various times surrounding the period of postnatal ethanol vulnerability. Ethanol selectively decreased Purkinje cell expression of TrkB and TrkC receptors following exposures within the vulnerable period (PN4-6). These results suggest that ethanol may induce loss of Purkinje cells by alteration of neurotrophic regulation at this critical stage.

Lack of Feminization of the Cremaster Nucleus in Cryptorchid Androgen Insensitive Rats

Androgens may control rat testicular descent via effects on the genitofemoral nerve or cranial gonadal ligaments. Androgen-mediated release of calcitonin gene-related peptide from the genito-femoral nerve (whose motoneuron cell bodies reside in the sexually dimorphic cremaster nucleus) may stimulate cremaster sac formation and testicular descent. Alternatively, androgens may cause regression of cranial gonadal ligaments and thereby allow the testes to descend. To evaluate these theories testicular position, and the cremaster sac and nucleus were studied in Tfm (androgen insensitive) rats. Testes were abdominal, inguinal and scrotal in 20%, 67% and 13% of Tfm male rats, respectively, and cranial ligaments were present in all cases. Mean cremaster nucleus motoneuron number was lower in female rats (70 +/- 14) but not significantly different between normal male (256 +/- 44) and Tfm male (231 +/- 42) rats, and it correlated poorly with testicular position. Calcitonin gene-related peptide immunoreactivity was rarely observed in cremaster motoneurons. These data suggest that the cremaster nucleus is not androgen-dependent, calcitonin gene-related peptide release from cremaster motoneurons is not the likely mechanism of testicular descent and persistent cranial ligaments may cause cryptorchidism in the Tfm rat.

A sexually dimorphic population of galanin-like neurons in the rat lumbar spinal cord : functional implications

The rat lumbar spinal cord contains a population of galanin- and cholecystokinin-containing neurons which are located dorsolateral to the central canal and project to the thalamus. New data are presented herein which reveal that the number of these neurons, as shown by galanin-like immunostaining, is sexually dimorphic with males containing 62% more of these neurons than females. This is the first demonstration of a sexually dimorphic population of intraspinal neurons which projects to higher CNS centers rather than to peripheral targets.

An in situ hybridization and immunohistochemical study of vasotocin neurons in the hypothalamus of water-deprived chickens

The distribution of immunoreactive vasotocin (IR-AVT) and AVT mRNA in the hypothalamus of White Leghorn cocks was determined by immunohistochemistry and in situ hybridization, respectively. In control birds that were provided with water ad lib, AVT mRNA was distributed in the periventricular and lateral regions of the hypothalamus in clusters of neurons that correspond structurally with the mammalian paraventricular (PVN) and supraoptic (SON) nuclei. Although the distribution of AVT, identified by immunohistochemistry of adjacent serial sections within the hypothalamus, was similar to the distribution of AVT mRNA, the possibility that some positive staining was due to mesotocin neurons was not excluded. Water deprivation for 2 and 4 days resulted in both an increase in levels of AVT mRNA per neuron and the number of AVT mRNA-containing cells. Additionally, water deprivation resulted in a decrease in the amount of IR-AVT per neuron. The results indicate that osmotic stimulation increases AVT gene expression not only in individual neurons but also by activating subpopulation of neurons that are not observed in normally hydrated birds.

Lack of feminization of the cremaster nucleus by prenatal flutamide administration in the rat and pig

PURPOSE The sexually dimorphic cremaster nucleus contains motoneurons that project via the genitofemoral nerve and theoretically direct androgen dependent testicular descent. The effects of flutamide on descent and masculinization of the cremaster nucleus were studied in the rat and pig. MATERIALS AND METHODS Flutamide was given to pregnant rats and pigs on days 16 to 22 and 65 to 113 of gestation, respectively. Tissues were perfused and examined at birth (pigs) or at age 30 days (rats). Spinal cords were removed, sectioned and immunohistochemically stained for serotonin (rats) or substance P (pigs) to demarcate the position of the cremaster nucleus and allow the determination of cremaster motoneuron number. RESULTS After exposure to flutamide testes were undescended in 6 of 9 rats and 7 of 10 pigs. Cremaster motoneuron number per nucleus were 288 +/- 22 in control versus 250 +/- 27 in flutamide treated rats, and 165 +/- 28 in control versus 148 +/- 24 in flutamide treated pigs. The decrease in motoneuron number by flutamide was significant in both species (p < 0.02) but it did not approach the levels in female rats (93 +/- 11) and pigs (57 +/- 12). Cremaster motoneuron number did not correlate with testicular position. Porcine undescended testes were associated with a significant increase in mean gubernacular volume. CONCLUSIONS Unlike other sexually dimorphic spinal cord nuclei masculinization of the cremaster nucleus appears to be largely androgen independent and it does not correlate with ipsilateral testicular descent. These data suggest that androgens do not mediate descent of the testes via the efferent limb of the genitofemoral nerve.

Walking a fine line: is it possible to remain an empathic physician and have a hardened heart?

Establishing an empathic physician–patient relationship is an essential physician skill. This chapter discusses the sexually dimorphic aspects of the neural components involved in affective and cognitive empathy, and examines why men and women medical students or physicians express different levels of empathy. Studies reveal levels of medical student affective or cognitive empathy can help reveal which medical specialty a student will enter. The data show students or physicians with higher empathy enter into specialties characterized by large amounts of patient contact and continuity of care; and individuals with lower levels of empathy desire specialties having little or no patient contact and little to no continuity of care. Burnout and stress can decrease the empathy physicians had when they first entered medical school to unacceptable levels. Conversely, having a too empathetic physician can let patient conditions and reactions interfere with the ability to provide effective care. By learning to blunt affective empathic responses, physicians establish a certain degree of empathic detachment with the patient in order to provide objective care. However, a physician must not become so detached and hardened that their conduct appears callous, because it is still important for physicians, especially those in specialties with a large amount of patient contact, to use empathic communication skills.

Galanin immunoreactivity in rat spinal lamina IX: Emphasis on sexually dimorphic regions

Fibers and puncta that contained galanin-like immunoreactivity (GAL-LI) were distributed within lamina IX in a heterogeneous fashion. In cervical spinal segments, GAL-LI was almost absent except for the phrenic nucleus, which received the most robust GAL-LI innervation in lamina IX. In high and mid-thoracic segments, GAL-LI was found in moderate amounts, but the number of GAL-LI fibers gradually diminished in a caudal fashion, so that in low thoracic segments GAL-LI was sparse. Throughout all thoracic segments, GAL-LI fibers surrounded some clusters of motoneurons, while other groups of motoneurons were devoid of GAL-LI fibers. In lumbar segments, three sexually dimorphic nuclei received sparse to moderate amounts of GAL-LI, while GAL-LI in the remainder of lumbar lamina IX was very sparse. In sacral spinal segments, GAL-LI was very sparse. These data indicate that fibers and puncta that contain GAL-LI preferentially surround motoneurons that innervate muscles associated with the axial skeleton, while motoneurons that innervate appendicular or tail-associated skeletal muscles only have an occasional GAL-LI fiber associated with them.

Cholecystokinin-8-like immunoreactivity is sexually dimorphic in a midline population of rat lumbar neurons

The gray matter surrounding the central canal of rat lumbar spinal cord contains a population of spinothalamic neurons which has been shown to co-contain cholecystokinin-8 (CCK-8) and galanin (GAL) (Ju, G., Melander, T., Hökfelt, T. and Frey, P., Immunohistochemical evidence for a spinothalamic pathway co-containing cholecystokinin- and galanin-like immunoreactivities in the rat. Neuroscience, 20 (1987) 439-456). A previous study from our laboratory has shown that female rats have only 62% the number of GAL-containing midline neurons present in males. Counts of CCK-8-containing midline neurons reveal that females have 59% the number observed in males. These results indicate that the degree of sexual dimorphism seen for CCK-8 is the same for GAL in these midline lumbar neurons, and suggests that androgens modulate the production of the co-contained GAL and CCK-8 in a similar fashion within male rat spinal cords.

Peptidergic innervation of the cremaster nucleus. I. A sexually dimorphic population of substance P-containing intraspinal neurons exists in the substance P pathway to the rat cremaster nucleus

The cremaster nucleus (CN) lies in the lumbar spinal cord and is sexually dimorphic: the male CN contains three times as many motoneurons as the female. The substance P (SP) innervation of the CN is also sexually dimorphic with males receiving a very prominent innervation which is greatly diminished in females. These investigations examined SP-containing neurons located in the ventral half of lamina IV and the lateral aspects of laminae V, VII, and IX, in lumbar spinal levels 1,2. SP-containing intraspinal neurons in these laminae are at least three times as numerous in males than females. This provides the first demonstration of a sexually dimorphic population of spinal neurons which is not motor or preganglionic in nature. These SP-containing interneurons are found within, or adjacent to, the SP-containing fibers which constitute the massive SP pathway to the male CN. Processes of these SP-containing neurons were observed to contribute to the formation of the SP pathway to the male CN. The immunohistochemically demonstrable presence of these lumbar 1,2, laminae IV-IX, SP-containing neurons validates former studies which suggested their existence (Gibson et al., Brain Research, 301 (1984) 243-251; Uda et al., Neurosci. Lett., 57 (1985) 185-190).

Androgens regulate the sexually dimorphic production of co-contained galanin and cholecystokinin in lumbar laminae VII and X neurons

A population of rat lumbar laminae VII and X putative spinothalamic (STT) neurons that co-contain cholecystokinin-8 (CCK) and galanin (GAL) are sexually dimorphic. Males have a significantly greater number of these neurons, as well as having greater optical densities for both neuropeptides than females. Optical densities for GAL and CCK immunoreactivities in these lumbar neurons in rats that have the testicular feminization mutation (Tfm) are not significantly different from females; however, the number of these lumbar neurons in Tfm rats is significantly smaller than in females. These data suggest that androgens, as well as functional androgen receptors (that Tfm rats lack), are necessary for the establishment of these sexual dimorphisms. Functionally, these CCK- and GAL-containing neurons in the deep lumbar laminae may contribute to the establishment of known sex differences in the affective component of somatic and visceral nociception, as well as the sexually dimorphic nature of some pelvic diseases, e.g., irritable bowel syndrome or cystitis.