The Association Between Prenatal Maternal Stress and Adolescent Affective Outcomes is Mediated by Childhood Maltreatment and Adolescent Behavioral Inhibition System Sensitivity.

Prenatal maternal stress is linked to offspring outcomes; however, there is little research on adolescents, behavioral, transdiagnostic outcomes, or the mechanisms through which relations operate. We examined, in N = 268 adolescents (Mage = 15.31 years; SD = 1.063; 57.8% boys) whether prenatal maternal stress is associated with adolescent affective outcomes; whether this association is mediated, serially, by childhood home atmosphere and adolescent behavioral inhibition system (BIS) sensitivity; and whether mediational effects are moderated by adolescent attention-deficit/hyperactivity disorder or maternal internalizing symptomology. Prenatal maternal daily stress and major life events were associated with adolescent outcomes through childhood negative atmosphere/neglect and BIS sensitivity, with no evidence of moderation. Results have implications regarding the effect of prenatal maternal stress on offspring outcomes and regarding corresponding sensitive periods.

Nerve stretch injury induced pain pattern and changes in sensory ganglia in a clinically relevant model of limb-lengthening in rabbits.

We used a model of tibial lengthening in rabbits to study the postoperative pain pattern during limb-lengthening and morphological changes in the dorsal root ganglia (DRG), including alteration of substance P (SP) expression. Four groups of animals (naive; OG: osteotomized only group; SDG/FDG: slow/fast distraction groups, with 1 mm/3 mm lengthening a day, respectively) were used. Signs of increasing postoperative pain were detected until the 10(th) postoperative day in OG/SDG/FDG, then they decreased in OG but remained higher in SDG/FDG until the distraction finished, suggesting that the pain response is based mainly on surgical trauma until the 10(th) day, while the lengthening extended its duration and increased its intensity. The only morphological change observed in the DRGs was the presence of large vacuoles in some large neurons of OG/SDG/FDG. Cell size analysis of the S1 DRGs showed no cell loss in any of the three groups; a significant increase in the number of SP-positive large DRG cells in the OG; and a significant decrease in the number of SP-immunoreactive small DRG neurons in the SDG/FDG. Faster and larger distraction resulted in more severe signs of pain sensation, and further reduced the number of SP-positive small cells, compared to slow distraction.

Preterminal and terminal axon arborizations in the substantia gelatinosa of cat's spinal cord.

Extensive terminal branchings of fine fibers in the substantia gelatinosa of Golgi-Kopsch preparations of the adult cat spinal cord were subjected to a semi-quantitative analysis. transverse sections suggest that these fibers are probably unmyelinated primary afferent elements of dorsal root origin. In transverse sections these elements pass medially and ventrally and shortly disappear due to a change in orientation. Similar thin fibers in sagittal sections can be followed for several hundred microns as they give rise to side branches that also run mainly in a longitudinal direction. The side branches divide in turn to produce preterminal axon arborizations. The arborizations were distributed in 150 mum wide zones in the dorsal horn region corresponding to Rexed's lamina II. The end terminals are large bulbs, usually preceded by two to three equally large en passant enlargements. Seven to eight terminals stem from each side branch. The terminals and enlargements are arranged in narrow (16-26 mum thick) sagittal sheets. The terminals of several side branches often converge upon a common region so that clusters of terminals occur within the sagittal sheet. It is proposed that these observations are consistent with the substantia gelatinosa (lamina II) as the termination of unmyelinated (C) primary afferent fibers and that the latter are the only type of primary fibers ending in this portion of the spinal cord.

Distribution of primary afferent fibers within the sacrococcygeal dorsal horn: an autoradiographic study.

The spinal segmental distribution and intersegmental course of primary afferent fibers were studied by injecting (by pressure or iontophoresis) tritiated amino acids (L-proline or L-leucine) into spinal ganglia of coccygeal and sacral segments and autoradiographically analyzing histological sections of the spinal cord, particularly those regions lying dorsal to the central canal. The results from eight cats and three monkeys are described. A heavy projection of primary afferent fibers to the marginal zone (lamina I), the substantia gelatinosa Rolandi (lamina II), and throughout the nucleus proprius (laminae III-IV) was demonstrated. The projections to these three areas appeared to be substantially independent. Primary afferent fibers were found to course rostrally and caudally within the marginal zone, in the midline dorsal to the central canal, in Lissauer's tract, and in the dorsal columns. A crossed projection passed by way of the dorsal commissure to the contralateral marginal zone and to a region ventrolateral to the contralateral nucleus proprius. There was a distinct medial-to-lateral shift in the termination of primary afferent fibers in the substantia gelatinosa and in the dorsal portion of the nucleus proprius. The most medial distribution occurred immediately caudal to the entry zone of the primary afferent fibers, and the most lateral at the cephalad end of the segment immediately rostral to the entry level. Small (iontophoretic) injections revealed circumscribed fields of termination, approximately 40 micrometers by 70 micrometers (dorsoventrally) by 400 micrometers or more (rostrocaudally) in the substantia gelatinosa.

Synaptic complexes formed by functionally defined primary afferent units with fine myelinated fibers.

The individual fine myelinated fibers of cutaneous mechanical nociceptors and "D-hair" receptors were identified by electrophysiological recording with micropipette electrodes in cats and monkeys. Their intraspinal terminations were labeled by iontophoresing horseradish peroxidase intracellularly and subsequent diaminobenzidine histochemistry. These terminations were examined with light and electron microscopy to determine the nature and organization of their synaptic contacts. Myelinated fibers of the mechanical nociceptors became unmyelinated before exhibiting many enlargements that made multiple synaptic contacts in the marginal zone (lamina I) and lamina V. Pre- or postsynaptic contacts were found only on enlargements. In the marginal zone of the cat, enlargements made simple axodendritic contacts or were scalloped, central terminals in glomeruli. In glomeruli, myelinated mechanical nociceptor enlargements were presynaptic to several dendritic appendages and postsynaptic to two different types of profiles. One type was interpreted as a presynaptic axon terminal, the other as a presynaptic, vesicle-containing, dendritic appendage. In lamina V of the cat the nociceptor synaptic complexes were similar, but simpler, and only axonal profiles were found to be presynaptic to them. In the monkey marginal zone and deep nucleus proprius, myelinated nociceptor terminations formed the central element of glomeruli, which consisted of postsynaptic dendritic appendages and presynaptic axon terminals. D-hair axons terminated in large numbers of enlargements in the nucleus proprius (laminae III and IV) and inner substantia gelatinosa (lamina IIi). Their large rounded enlargements formed the central terminals in glomeruli and were presynaptic to both ordinary and vesicle-containing dendritic appendages; the presynaptic dendritic profiles also often contacted each other. Profiles interpreted as axonal in origin were the only terminals presynaptic to the primary ending within the D-hair glomeruli. The results suggest that transfer of primary afferent information occurs only at enlargements of the primary fiber and that each primary fiber enters into more than one kind of synaptic arrangement. They also point out that synaptic glomeruli are common to functionally different types of primary afferent fibers and that the internal organization of glomeruli varies with the kind of primary fiber and the locus of the complex.

Dendritic arborization and axon trajectory of neurons in the hypothalamic arcuate nucleus of the rat--updated.

Neurons in the hypothalamic arcuate nucleus (arcuate neurons) were traced on Golgi-impregnated sections. Dendrites of arcuate neurons showed characteristic orientation patterns. Dendrites along the lateral side follow the convex border of the nucleus by running parallel to the tanycyte processes. Neurons located in the ventrolateral portion of the nucleus have dendrites running parallel to the basal surface of the hypothalamus. Fine, beaded axons of arcuate neurons project mostly ventrally, and less frequently dorsally and dorsolaterally. Ventrally projecting axons converge towards the tuberoinfundibular sulcus which emerges into the ventral portion of the arcuate nucleus from below.

Altered distribution of dorsal root fibers in the rat following neonatal capsaicin treatment.

Distribution of primary afferent fibers was studied in intact and neonatally capsaicin treated rats by the application of horseradish peroxidase to the central branch of the transected lumbar dorsal roots. Coarse primary afferent fibers entered the spinal cord through the larger medial portion of the rootlet and arborized in the deeper part of the dorsal horn (laminae III and IV). Fine fibers reached the spinal cord through the smaller lateral portion of the rootlet and arborized in the superficial portion of the dorsal horn (lamina I and outer portion of lamina II). The technique used was inadequate to stain fine, unmyelinated primary afferent fibers terminating in the larger inner portion of lamina II. After neonatal capsaicin treatment (50 mg/kg) the flame-shaped arborizations of thick primary afferent fibers terminating in intact rat in laminae III and IV spread dorsally and occupied the inner portion of lamina II in the larger lateral sector of the dorsal horn. Medially the dense arborization of a different type of thick primary afferent fibers sprouted up to the white-gray border. The border between the lateral and medial sector was sharp and only slightly varied in localization from experiment to experiment. The sprouting fibers established complicated synaptic contacts with dendrites and axon terminals. The rearrangement of primary afferent fibers after neonatal capsaicin treatment confirmed earlier results and revealed a mediolateral difference in the fiber organization of the dorsal horn indicating differences in the projection from hairy vs non-hairy skin areas.

Distribution of neurons expressing calcitonin gene-related peptide mRNAs in the brain stem, spinal cord and dorsal root ganglia of rat and guinea-pig.

In situ hybridization histochemistry was used to localize calcitonin gene-related peptide mRNAs in spinal cord, brain stem and dorsal root ganglion neurons of the rat and guinea-pig. A 32P-labeled 23-base-long (23mer) oligodeoxyribonucleotide (oligomer) complementary to calcitonin gene-related peptide mRNA sequences encoding residues 23-30 of calcitonin gene-related peptide was used primarily as a probe (CGRP I probe). A 32mer complementary to mRNA sequences for residues 10-20 of calcitonin gene-related peptide (CGRP II probe) was also used as a positive control for specificity of the 23mer for calcitonin gene-related peptide mRNA. In both the guinea-pig and rat calcitonin gene-related peptide mRNA was localized specifically to neurons of the dorsal root ganglion, to spinal motoneurons and to motoneurons of the hypoglossal, facial and accessory facial motor nuclei. Differences in the distribution of calcitonin gene-related peptide mRNA between the rat and guinea-pig included a higher proportion of rat dorsal root ganglion neurons containing calcitonin gene-related peptide mRNA and the localization of calcitonin gene-related peptide mRNA to motoneurons of the ambiguus motor nucleus, parabrachial and peripeduncular nucleus of the rat but not the guinea-pig. In the guinea-pig, in contrast, calcitonin gene-related peptide mRNA was localized also to motoneurons of the abducens, trigeminal, trochlear and oculomotor nerves. The neuronal groups in the intact rat found here to contain calcitonin gene-related mRNA have also been shown previously to contain calcitonin gene-related peptide immunoreactivity in colchicine-treated rats. Colchicine-treated rats, however, have been found to contain additional groups of calcitonin gene-related peptide immunoreactive neurons which, in the intact rats used in the present study, showed no detectable hybridization with the calcitonin gene-related peptide probe.

Morphological properties of physiologically characterized lamina III neurones in the cat spinal cord.

Six lamina III interneurones of the cat spinal cord were impaled and stained with intracellular injections of horseradish peroxidase. The responses of these neurones varied considerably: 1 neurone responded to light brushing of its receptive field, whilst 4 cells were excited by strong pressure. Morphologically, they were also a heterogeneous population. Two neurones had rostro-caudally orientated dendritic arbors that were confined to the lamina, while four of the cells were vertically orientated and possessed dendrites that crossed lamina boundaries. There was no correlation between the physiological characteristics of a neurone and its morphology. Three of the vertically orientated neurones were examined ultrastructurally. The first of these cells received several types of synaptic terminal which were distributed in an organised pattern over the entire dendritic tree. This neurone possessed recurrent dendrites which participated in a variety of complex synaptic arrangements. The second neurone also participated in a variety of synaptic arrangements, including glomeruli in lamina II, and received contacts from vesicle-containing dendrites. It gave rise to collateral axons which arborized in lamina II and formed boutons which formed synapses with dendrites. The third cell possessed varicose dendrites which were probably artifactual. It is concluded that lamina III interneurones are a heterogeneous population by electrophysiological, morphological and ultrastructural criteria. They differ in many respects from lamina II neurones and from the cells of origin of ascending systems. The diversity of their inputs and their variation in morphology suggests that they receive input from a variety of primary afferent fibres and dorsal horn neurones and hence may integrate information from these sources.

Colchicine enhances mRNAs encoding the precursor of calcitonin gene-related peptide in brainstem motoneurons.

Hybridization signals indicating mRNAs encoding the precursor of calcitonin gene-related peptide (CGRP) and CGRP immunoreactivity were detected on parallel sections containing brainstem motor nuclei using in situ hybridization histochemistry and immunohistochemistry. In untreated and saline-injected rats the motoneurons in the hypoglossal, facial motor nuclei and in the ambiguus nucleus showed weak to moderate hybridization signals. In these motoneurons CGRP immunoreactivity was restricted to the Nissl bodies of the perikarya. Twenty-four and 42 hours after intracerebroventricular colchicine injection the intensity of both the hybridization signal and the immunoreaction product increased. The distribution of CGRP immunoreactivity changed from discrete perikaryal localization to diffuse reaction in the perikarya and along the proximal dendritic tree. Motoneurons in the rest of the brainstem motor nuclei (VIth, Vth, IVth and IIIrd) of untreated and saline-injected rats showed neither hybridization signal nor CGRP immunoreactivity. After intracerebroventricular injection of colchicine these motoneurons showed both hybridization signal and CGRP immunoreactivity. In all nuclei the size of motoneurons decreased and their Nissl structure changed to an amorphous basophilic mass following colchicine treatment.

Species-specific expression of cholecystokinin messenger RNA in rodent dorsal root ganglia.

The expression of cholecystokinin (CCK) messenger RNA (mRNA) was examined in dorsal root ganglia of rat and guinea pig using in situ hybridization histochemistry and RNA (Northern) blot hybridization with synthetic oligodeoxyribonucleotide (oligomer) probes. In guinea pig, CCK mRNA was detected in small and medium-sized neuronal perikarya comprising approximately 10-15% of the total dorsal root ganglia cell population. In contrast, in neurons of rat dorsal root ganglia, CCK mRNA was not detectable. Northern blot analyses revealed a single CCK mRNA species of expected size (0.8 kb) in guinea pig, but not rat, dorsal root ganglia. A 0.8 kb CCK mRNA was, however, detected in cortex of both rat and guinea pig. These data suggest that CCK is normally not synthesized in neurons of rat dorsal root ganglia and that there are species differences in CCK gene expression in mammalian sensory ganglia.

Semi-quantitative analysis of somatostatin mRNA distribution in the rat central nervous system using in situ hybridization.

The distribution of somatostatin mRNA in the rat brain has been examined by in situ hybridization using 32P-labelled oligonucleotide probes. Numerous telencephalic and diencephalic areas contained labelled cells with the largest numbers of cells occurring in the anterior olfactory nucleus, olfactory and entorhinal cortices, hippocampus, neocortex, caudate nucleus, accumbens, septum, amygdala and periventricular nucleus. Fewer labelled cells occurred in the mesencephalon and rhombencephalon but groups were seen in the region of the central grey, lateral lemniscus, parabrachial and tegmental nuclei, medial longitudinal fasciculus and nucleus of the solitary tract. This distribution closely matches published maps of the distribution of somatostatin-immunoreactive cell bodies. The intensity of individual cell labelling has also been quantified using image analysis and compared with the intensity of somatostatin immunocytochemical cell staining. In situ hybridization cell labelling varied both within different regions and from region to region. Highest labelling was seen in the periventricular nucleus of the hypothalamus followed by telencephalic regions such as cortex, hippocampus and the medial nucleus of the amygdala. In contrast all brainstem areas had low levels of labelling with the lowest levels of the brain occurring in the dorsolateral tegmental nucleus. Somatostatin immunocytochemistry showed similar variations such that the intensity of cell immunostaining broadly paralleled the intensity of cell in situ hybridization labelling. Thus both peptide and mRNA levels were much lower in brainstem cells than in forebrain, although a close correlation between immunocytochemistry and in situ hybridization was not seen in all brain regions.

Approximation of missing sections in computer reconstruction of serial EM pictures.

Replacing missing contours or completing a series of contours of biological structures has been investigated. The paper reports on a method for approximating lateral surfaces between adjacent contours and generating new artificial outlines by slicing the interpolated surface parallel with the original sections. The research method was developed mainly due to the need to fill in damaged or missing ultrathin sections in longer series, since their absence may impede the three-dimensional visualization of selected components. The new program attached to the shape reconstruction program system existing at Second Department of Anatomy, Semmelweis University Medical School, Budapest, Hungary also permits new contours to be generated into the models of single or branching objects. Besides replacing missing contours, the program makes the reconstructed model more precise.

Diffusional barrier around the hypothalamic arcuate nucleus in the rat.

The contour lines of horseradish peroxidase injection sites in the ventrobasal hypothalamus were distorted by the border between arcuate and ventromedial nuclei as well as between arcuate nucleus and median eminence. The dense array of tanycyte processes is assumed to isolate the arcuate nucleus from the neighboring territories by establishing a diffusional barrier surface.

Quantitative analysis of dendritic protrusions in the medial preoptic area during postnatal development.

Dendritic architecture and distribution of dendritic protrusions were studied on Golgi-impregnated neurons in the medial preoptic area (MPOA) of infant rats (7 and 20 days old), of animals at puberty (34 days old) and of postpubertal rats (90 days old) using computerized image analysis. The protrusions showed a peak distribution on the proximal portion of the dendritic tree in infant rats. At puberty and in postpubertal animals protrusions disappeared almost completely along the most proximal portion of the dendritic tree. The data suggest that differentiation of certain MPOA neurons is still in progress at puberty.

Is the knife-cut in the hypothalamus a permanent barrier to regrowth of nerve fibers? -an affirmative answer.

The regenerative capacity of nerve fibers was studied in adult female rats. Horseradish peroxidase was injected into the anterior hypothalamus lateral to the suprachiasmatic nucleus 4 days, 6 weeks and 4 months, respectively, following an archiform retrochiasmatic knife-cut. The trajectory of the stained fibers was examined on horizontal sections of the hypothalamus. No nerve fibers could be seen sprouting across the scar-tissue of the knife-cut regardless of the survival time. In one rat (6-week survival time) a bundle of fine nerve fibers turned in a medial direction at the caudal end of the knife-cut, suggesting that sprouting fibers were destined to reinnervate parts of the deafferented medial-basal hypothalamus.

Synapses upon the axon origin of dorsal horn neurons in the rat spinal cord.

Axon terminals synapsing with axon hillocks or origins of Golgi-impregnated and gold-toned neurons in the dorsal horn of the rat were shown in serial electron micrographs. Synapses occurred irrespective of the site (perikaryon or dendrite) and mode (with or without an axon hillock) of the axon origin. The synapsing axon terminals contained 3 populations of vesicles: pleomorphic and flattened synaptic vesicles and a combination of pleomorphic and dense-core vesicles. The membrane thickening in the axon-axon hillock synapses was of the symmetrical type.

Sensitivity of the short-range spinal interneurons of the cat to experimental spinal cord trauma.

The technique of retrograde axoplasmic transport was used to demonstrate the effect of experimental spinal cord injury on the spinal interneurons in the upper lumbar and lower thoracic segments of cats. Force of varied intensity was applied to the dorsal surface of the spinal cord and horseradish peroxidase (HRP) was injected into the next caudal segment. A large impact (250 to 350 gm-cm) inducing permanent paraplegia of the hind legs blocked the axoplasmic transport instantaneously in both cranial and caudal directions. If 1 week elapsed between the trauma and injection, neurons cranial to the trauma did not show any evidence for retrograde axoplasmic transport, while few neurons in the caudal direction were labeled with HRP. A moderate impact (150 gm-cm) which rendered the animals only transiently paraplegic spared the axoplasmic transport in some neurons both cranially and caudally to the injection. No obvious recovery or additional loss in the number of HRP-labeled neurons could be found in the cats if the injections followed the trauma by 1 week. The loss of spinal cord neurons following the injury seems to be the immediate mechanical consequence of the trauma.

Position and size of the axon hillock in various groups of neurons.

The origin of the axon was studied in Golgi-Kopsch impregnated specimens prepared from the spinal cord and brain of adult rats. Five types of neurons were sampled: large ventral horn neurons, neurons in the intermediate zone and ventral horn of the spinal cord, antenna-type neurons in the spinal dorsal horn, neurons in the thalamus, and neurons in the hypothalamus. The axon originated from the perikaryon in 76% of the large ventral horn neurons and in 64% of the neurons in the thalamus. In contrast, the axon emerged from one of the dendrites in 75% of the neurons in the intermediate zone and the ventral horn of the spinal cord and in 68% of the neurons in the hypothalamus. In the case of the antenna-type neurons in the spinal dorsal horn, the axon often originated from one of the dendrites, but never from a dorsally oriented dendrite. The mean distance of the axon hillock of dendritic origin was the longest in the neurons in the intermediate zone and the ventral horn of the spinal cord. The size of the axon hillock was proportional to the size of the perikaryon. The impregnated portion of the axon was longest in the large ventral horn neurons.