A case of insertional translocation involving chromosomes 2 and 4.

We report on a 6-year-old Caucasian boy with direct insertion of genetic material from the short arm of chromosome 4 to the short arm of chromosome 2. He was referred for evaluation because of global developmental delay and seizure disorder. A karyotype performed at 4 1/2 months of age, by a laboratory elsewhere, reportedly showed a deletion of chromosome 4(p12). When we saw him, he had macrocephaly, hypotonia, psychomotor retardation, multiple minor congenital anomalies, and EEG abnormalities. Repeat chromosomes performed by our laboratory revealed that his karyotype was 46,XY,dir ins(2;4)(p24;p15.3p13). Fluorescence in situ hybridization (FISH) analysis, using chromosomes 2 and 4 painting probes confirmed that material from 4p has been translocated to 2p. Also, FISH analysis using the Wolf-Hirschhorn critical region probe revealed that both loci are intact. Parental chromosomes were normal. This complex rearrangement, though it appears balanced, probably might have resulted in either a structural loss of genetic material or functional loss of a gene action. Thus, his phenotype could be explained by this de novo insertion of chromosome 4 material into chromosome 2. There is no reported case of this specific chromosome rearrangement.

Spinal neurogenesis and axon projection: a correlative study in the rat.

The purpose of the present study was to determine the relationship between the duration of a spinal neuron's neurogenic period and the length of its axon or level of projection. Spinal segment L1 was chosen for examination and neurons were divided into four projection groups: 1) supraspinal projection (SSp), 2) long ascending propriospinal (LAPr), 3) short ascending propriospinal (SAPr), and 4) descending propriospinal (DPr). To determine the duration of the neurogenic period for each group, 3H-thymidine was administered to fetal rats during the proliferative period for spinal neuroblasts on one of embryonic (E) days E13 through E16. Between 50 and 100 days after birth neurons in each group were labeled with the retrograde fluorescent tracer Fluoro-Gold. To demonstrate nerve cells with SSp projections, spinal cords were hemisected at spinal segment C3 in one group of animals and Fluoro-Gold was applied to the sectioned surface of the cord. Three additional sets of animals were used to label nerve cells with LAPr, SAPr, and DPr projections by injecting Fluoro-Gold into the gray matter at spinal segments C6, T12, and L5, respectively. Neurons labeled with both Fluoro-Gold and 3H-thymidine and neurons labeled with Fluoro-Gold alone in each animal in each group were counted and the data statistically analyzed. Results showed that within each spinal lamina neurons with different projections were generated, i.e., completed cell division, at significantly different rates. Neurons with the longest axons, those with SSP projections, were generated first. These were followed by those with LAPr projections, and finally those with SAPr and DPr projections. In most laminate there was no significant difference between the neurogenic periods of rostrally projecting short propriospinal (SAPr) neurons versus caudally projecting short propriospinal (DPr) neurons. It was concluded that the duration of the neurogenic period for a given group of neurons within each spinal lamina is inversely related to the distance between the nerve cell and its projection site regardless of the direction of its projection.

Neurogenesis of ascending supraspinal projection neurons: ipsi- versus contralateral projections.

The present study tests the hypothesis that contralaterally projecting supraspinal projection neurons (SPNs) are generated prior to ipsilaterally projecting SPNs. Neuronal time of origin was determined by injecting pregnant rats with tritiated thymidine on one of embryonic (E) days E12 through E15. In mature offspring of thymidine-treated dams, SPNs in the lumbar cord were retrogradely labelled with True Blue delivered at the site of a hemisection in spinal segment C3. Ipsi and contralaterally projecting SPNs in laminae I, VII and VIII and the lateral spinal nucleus, which are known to give rise to long sensory pathways, were generated simultaneously throughout their neurogenic period (E12-E14), while ipsilaterally projecting SPNs in lamina IV and the nucleus dorsalis, which give rise to short sensory pathways, completed neurogenesis one day later (E15). Results suggest that the projection target and its distance from the nerve cell body of origin are more consistent correlates of the duration of the neurogenic period than the course of the axon.

Neurogenic period of ascending tract neurons in the upper lumbar spinal cord of the rat.

Although the neurogenic period for neurons in the lumbar spinal cord has been clearly established (Days 12 through 16 of gestation), it is not known when the neurogenesis of ascending tract neurons is completed within this period. The purpose of the present study was to determine the duration of the neurogenic period for projection neurons of the ascending tracts. To label neurons undergoing mitosis during this period, tritiated thymidine was administered to fetal rats on Embryonic (E) Days E13 through E16 of gestation. Ascending tract neurons of the lumbar cord were later (Postnatal Days 40-50) labeled in each animal with a retrograde tracer, Fluoro-Gold, applied at the site of a hemisection at spinal cord segment C3. Ascending tract neurons which were undergoing mitosis in the upper lumbar cord were double labeled, i.e., labeled with both tritiated thymidine and Fluoro-Gold. On Day E13, 89-92% of the ascending tract neurons were double labeled; on Day E14, 35-37%; and on Day E15, 1-4%. Results showed, then, that some ascending tract neurons were double labeled through Day E15 and were, therefore, proliferating in the final one-third of the neurogenic period. Ascending tract neurons proliferating on Day E15 were confined to laminae III, IV, V, and X and the nucleus dorsalis. Long tract neurons in the superficial dorsal horn (laminae I and II), on the other hand, were found to have completed neurogenesis on Day E14 of gestation. Results of the present study show that spinal neurogenesis of ascending projection neurons continues throughout most of the neurogenic period and does not completely follow the well-established ventral to dorsal gradient.

Nerve cell bodies in the dorsal funiculus of the rat spinal cord.

Nerve cell bodies located within the white matter of the dorsal funiculus (DF neurons) have been previously observed but not described in detail. The present study examines the morphology, ontogeny, and projection of DF neurons utilizing Fluoro-Gold as a retrograde tracer, alone, and in combination with tritiated thymidine autoradiography in the spinal cord of the rat. DF neurons were consistently labelled in spinal segments T13 through L2 following injections of Fluoro-Gold into the cerebellum. The cell bodies of DF neurons were small to medium in size, fusiform to multipolar in shape, and were located on the side ipsilateral to the injection site. Cell counts revealed approximately five labelled cells per millimeter along the longitudinal axis. An examination of neurogenesis using tritiated thymidine combined with Fluoro-Gold showed that DF neurons have relatively late birthdates as do other spinocerebellar neurons of the dorsal horn. Retrograde axon tracing studies in the spinal cord using Fluoro-Gold showed that DF neurons project rostrally via the ipsilateral lateral funiculus. The significance of the presence of nerve cells in the dorsal funiculus is unclear, but judging from their location, ontogeny, and projection, DF neurons are probably derived from the same pool of neurons as those in the Nucleus dorsalis.

Neurons with asymmetrical dendritic arbors in the substantia gelatinosa of the rat spinal cord.

Two types of neurons were observed in the substantia gelatinosa (SG) of the rat spinal cord which exhibit wide variations in dendritic symmetry. As demonstrated with the Golgi technique, "islet" cells with short dendritic arbors and "type III stalk" cells display dendritic patterns which vary from a bipolar type arrangement with two dendritic arbors of nearly equal dimensions to a unipolar arrangement with a dendritic arbor which extends in only one direction. Examination of the morphology and dendritic development of these neurons shows that they are unique compared with other SG neurons in that they have short, longitudinal dendritic arbors which undergo maturation relatively late in the postnatal period. As is discussed, variations in dendritic symmetry are probably dependent on the location of the terminal fields of primary and/or other types of afferents which are formed earlier in development.

A simple method for combining HRP-TMB histochemistry with tritiated thymidine autoradiography on the same tissue section.

In order to determine the sequence of development of various types of spinal neurons defined by their projection, we have developed a method for combining tritiated thymidine autoradiography for birthdate determination, with the demonstration of retrogradely transported horseradish peroxidase (HRP) using tetramethylbenzidine (TMB) as the chromogen. Because of its greater sensitivity, TMB is the chromogen of choice for the demonstration of HRP. However, the HRP-TMB reaction product is unstable and completely destroyed when the tissue is processed for autoradiography. The present study describes the use of osmium tetroxide as a postreaction stabilizing agent which preserves the HRP-TMB reaction product in the form of a dark brown precipitate which is not destroyed when the tissue is subsequently processed for autoradiography. Background levels of autoradiographs stabilized with the osmium procedure are extremely low.

Structure and development of central arborizations of hair follicle primary afferent fibers.

The present study describes the structure and development of the flame-shaped central arborizations of hair follicle (HF) afferents in the lumbosacral spinal cord of the rat. Tissue was processed according to the rapid Golgi method at successive stages of development from embryonic day 17 through postnatal day 30. Collaterals of most HF afferents were found to enter the dorsal horn via a characteristic U-shaped pathway which often parallels the vascular pattern. The HF collaterals can first be identified at embryonic day 19 and by postnatal day 5 they have established the dorso-ventral and rostro-caudal limits of their field of arborization. Dorsally the arbors extend no further than the inner zone of lamina II (IIi) at any stage of development. Short aberrant branches were observed on some HF collaterals during the prenatal period but none of these developed synaptic terminals or contributed branches to other arbors. Each HF collateral formed a single well defined flame-shaped arbor with a distribution and branching pattern which could be distinguished from that of other afferents throughout the postnatal developmental period. Two types of HF collaterals were observed. Structure and distribution patterns suggest that type I collaterals are derived from G and T HF afferents while collaterals in the type II category are probably derived from both G and T as well as D (A-delta) HF afferents. Type I collaterals divide into well defined medial and lateral collateral branches which arborize mainly in lamina III with a few branches to lamina IV. Type II collaterals are characterized by a delicate arbor which is more vertically oriented than type I arbors.(ABSTRACT TRUNCATED AT 250 WORDS)