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1

Sex differences in the number and morphometric properties of muscle spindles in the rat medial gastrocnemius muscle

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Gartych M., Bukowska D., Mierzejewska-Krzyzowska B., Celichowski J.
Acta Neurobiologiae Experimentalis
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2017
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tom 77
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nr Suppl.1
INTRODUCTION: There are considerable differences in a number and density of muscle spindles in various skeletal muscles. Considerable sex differences in muscle mass and diameter of extrafusal muscle fibres suggest that muscle spindle density in muscles and morphometric properties of spindles: the diameter, the number and diameters of intrafusal muscle fibres are also different in males and females. Similar number of γ‑motoneurons in male and female rats suggest similar number of muscle spindles but their lower density in males. AIM(S): The aim of the study was to check sex differences in the number, the density and morphometric properties of muscle spindles in rat medial gastrocnemius muscle. METHOD(S): Medial gastrocnemius muscles were excised from two male and two female three-month old Wistar rats. Muscles were stored in 4% formalin solution and then cut into 10 and 20 µm slices stained with methylene blue and magenta. The light microscopy (Nikon microscope with camera and NIS Elements program) was used to calculate a number of muscle spindles in the muscle. The morphometric properties of spindles were measured on the equatorial regions. RESULTS: The number of muscle spindles is similar in male (13), in female (13–14) muscles. However, the density of spindles was different: 86–92 in males and 51–57 mg of the muscle mass per one spindle in females. There were also slight differences in a number of intrafusal muscle fibres in one spindle (in males: 4.3; range 3–8 and in females: 4.5; range 2–7). The diameters of intrafusal fibres on the equatorial regions amounted to: 6.7±2.1 µm in males and 6.6±2.6 µm in females. However, it was noticed that diameters of the male muscle spindles are larger than in the female ones: 21.4±7.3 µm and 20.2±5.7 µm, respectively. CONCLUSIONS: The main sex difference concerns density of muscle spindles which is lower in males than in females.
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Divergent axons within olivocerebellar projection to the pyramis and uvula revealed by a retrograde double labeling technique in the rabbit

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr S
The study was initiated to determine the projection pattern of the climbing fibers from the inferior olivary complex (IO) to the two lobules of the caudal vermis, both independent and by way of axonal collaterals. Different fluorescent tracers, Fast Blue and Diamidino Yellow, unilaterally injected into the pyramis and uvula in the seven rabbits, resulted in single (n=32020) and double (n=403) labeling of neurons in defined restricted regions of the contralateral IO. These neurons, parent for independent and collateral projections, respectively, clustered in two aggregations (lateral and medial) through the rostrocaudal extent of IO. IO consists of a complex of three nuclei: the dorsal (DAO) and medial (MAO) accessory olive, and the principal olive (PO). MAO is accompanied by the dorsomedial cell column (dmcc) and the β nucleus (β). PO is composed of the dorsal (dlPO) and ventral (vlPO) laminas and comprises the ventrolateral outgrowth (vlo) and dorsal cap (dc). Distribution pattern of single labeled neurons indicates that the strongest projection to the pyramis and uvula arises from the lateral region of entire extent of MAO and that to the pyramis is more intensive. Neurons in the lateral region of DAO send numerous fibers exclusively to the pyramis. The rostral part of β more frequently supplies the uvula whereas the caudal part - the pyramis. Neurons of dmcc in similar degree project to the two cerebellar targets, however, from the rostral part projection is stronger to the uvula. Weak connections come from vlo and vlPO, and from dc only to the uvula. Neurons participating in collateral projection were found in the lateral region of MAO, the entire extent of β, the rostral and caudal levels of dmcc, and in vlo. To sum up, (1) the IO-pyramis and IO-uvula projecting neurons vary in number and in distribution, and (2) there is a small population of the IO neurons which project by divergent axons.
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The number and motor innervation of muscle spindles in the medial gastrocnemius of male and female rats

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Celichowski J., Gartych M., Mierzejewska-Krzyzowska B., Bukowska D.
Acta Neurobiologiae Experimentalis
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2019
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tom 79
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nr Suppl.1
INTRODUCTION: There are numerous sex differences concerning male and female skeletal muscles, including muscle mass, the number and diameter of muscle fibers, and number of motor units; however, there is no data concerning the number and density of muscle spindles, which are the most important muscle proprioceptors. AIM(S): The experiments aimed to determine the number of gamma motoneurons, number and density of muscle spindles, as well as, their morphometric properties for rat medial gastrocnemius. METHOD(S): The motoneurons were stained with the horseradish peroxidase and neurons exceeding 27.5 µm were accepted as gamma size. Muscle spindles were counted in muscle cut into 5‑10 µm sections. The rat medial gastrocnemius was innervated by a similar number of gamma motoneurons (27.6±3.9 and 28.7±6.6, p>0.05) for female and males, respectively. RESULTS: However, size of gamma motoneurons was higher in males (23.5±2.9 µm) than in females (21.6±2.9 µm) (p<0.01). The number of muscle spindles amounted to 13.16±1.25 and 14.0±2.71 for female and males, respectively (p>0.05). The diameter of intrafusal muscle fibers was similar in males (5.16±2.43 µm) and in females (5.37±2.27 µm) (p>0.05), whereas the number of intrafusal muscle fibers was smaller in females (4.76±1.23) than in males (5.2±1.89, p<0.05). The mass of studied muscle was 61% higher in males (1.08 g vs. 0.66 g in females); therefore, the muscle mass per one spindle was 77 mg in males and 50 mg in females. CONCLUSIONS: The number of muscle spindles as well as their motor innervation by gamma motoneurons is similar in male and female rats, but the density of muscle spindles is considerably higher in females. FINANCIAL SUPPORT: Supported by grant 2018/31/ B/NZ7/01028.
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Ultrasound elastography in clinical diagnostics and in scientific research on muscles

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Rosicka K., Arlet J., Bukowska D., Mierzejewska-Krzyzowska B.
Central European Journal of Sport Sciences and Medicine
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2019
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tom 27
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nr 3
5

Differences in the ponto-cerebellar projection to the cerebellar caudal vermis and hemisphere: a retrograde double labeling study in the rabbit

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Zguczynski L., Bukowska D., Mierzejewska-Krzyzowska B.
Acta Neurobiologiae Experimentalis
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2011
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tom 71
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nr S
Following paired unilateral injections of fluorescent tracers FB (Fast Blue) and DY (Diamidino Yellow) into the the rostral and caudal paramedian lobule (rPML, cPML) as well as the pyramis (Pr) and uvula (Uv), the distribution pattern of retrogradely labeled neurons in the pontine nuclei (PN) indicates that these two ponto-cerebellar projections are organized topographically. Both projections are bilateral. Projection to PML originates from the caudal two-thirds whereas that to the Pr and Uv - from the entire rostrocaudal extent of PN. Participation of individual PN nuclei in the projections differs. The strongest connections to rPML/cPML and Pr/Uv send the dorsolateral (44% and 56%, respectively) and paramedian (24% and 26%, respectively) nuclei. The involvement of the lateral (20% and 14%) and peduncular (12% and 4%) nuclei is weaker. The ventral pontine nucleus sends no fibers to PML or Pr and Uv. Moreover, following injections into the rPML and cPML, some regions where FB and DY single labeled neurons were intermingled, contained in addition small number of double labeled neurons. These neurons are parent for collaterals projections to both parts of PML. Differences in projections under study may arise from various functions of the PML (rPML and cPML receive afferent information from the forelimb and hindlimb, respectively) and the caudal vermal lobules (Pr receives spinal cord afferents related to innervation of axial and proximal-limb muscles, and Uv is interconnected with the vestibular nuclei)
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Differences in the ponto-cerebellar projection to the vermis: double labeling method in the rabbit

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Mierzejewska-Krzyzowska B., Bukowska D., Zguczynski L.
Acta Neurobiologiae Experimentalis
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2007
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tom 67
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nr 3
7

Topography in the inferior olive projections to the uvula and paramedian lobule in the rabbit

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B.
Acta Neurobiologiae Experimentalis
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2005
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tom 65
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nr 3
8

Evidence for diverging projection from the dorsal column nuclei to the cerebellar pyramis and uvula

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B.
Acta Neurobiologiae Experimentalis
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2007
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tom 67
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nr 3
9

The differences in organization of motor nucleus of the medial gastrocnemius muscle in male and female rat

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Mierzejewska-Krzyzowska B., Bukowska D., Taborowska M., Celichowski J.
Acta Neurobiologiae Experimentalis
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2013
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tom 73
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nr Suppl.1
The aim of this study was to recognize the sex differences in the architecture of the medial gastrocnemius (MG) motor nucleus in the same age rats. The retrogradely labeled motoneurons in the MG motor nucleus were studied following a bath of proximal stump of the transected MG nerve in the horseradish peroxidase. The rostrocaudal distribution of motoneurons along the spinal cord and on transversal sections as well as size and density of motoneurons in the motor nucleus were determined from serial microscopic images. It was shown that length of the motor nucleus in L4–L6 segments was 37% greater in males. Three types of motoneurons with different soma diameter were revealed: α one (27.5–40.0 μm), α two (greater than 40.0 μm) and γ (smaller than 27.5 μm). The density of α one and α two motoneurons was 15% higher in females. However, sex differences between the number of α one and α two motoneurons were more significant in group of α one motoneurons. The number of α one motoneurons in the motor nucleus was 24% higher in males than females (41 versus 33 motoneurons). The density of α motoneurons was 31% higher in females. It is concluded that length of the MG motor nucleus is greater in males, but the density of α and γ motoneurons was higher in females.
10

Pontocerebellar projection to the rabbit paramedian lobule by means of axonal collaterals: evidence for intralobular connections

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B.
Acta Neurobiologiae Experimentalis
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2003
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tom 63
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nr 4
11

Branching axon from pons and raphe nuclei project to cerebellar paramedian lobule as studied with the double fluorescent retrograde tracing technique

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Mierzejewska-Krzyzowska B., Sikora E., Zguczynski L.
Folia Morphologica
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1996
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tom 55
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nr 4
12

Collateral projections of trigeminal sensory neurons to both cerebellar paramedian lobules in the rabbit: demonstration by fluorescent double labeling study

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B., Sikora E.
Acta Neurobiologiae Experimentalis
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1998
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tom 58
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nr 4
13

Distribution and immunohistochemical characteristics of cerebellar projecting neurons in the locus corelueus complex of the rabbit

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Bukowska D., Zguczynski L., Mierzejewska-Krzyzowska B., Sienkiewicz W., Kaleczyc J.
Acta Neurobiologiae Experimentalis
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2015
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tom 75
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nr Supl.
BACKGROUNDANDAIMS: The locus coeruleus complex (LCx), subdivided into nucleus coeruleus proper (LC) and subcoeruleus (LCα) is the major noradrenergic nucleus of the brain. Only a few data concern  LCx-cerebellar projection. The aim of study was to identify LCx neurons projecting to the caudal vermis and determine their immunohistochemical characteristics. METHODS: The retrograde axonal transport method of fluorescent tracers Fast Blue (FB) and Diamidino Yellow injected respectively into lobules VIII and IX was used. In addition, double labeling immunofluorescence was applied to investigate the expression and coexistence of dopamine-β-hydroxylase (DBH) and choline acetyltransferase as well as neuropeptides including neuropeptide Y (NPY), somatostatin (SOM), leu-enkephalin (LENK) and substance P in LCx-cerebellar projecting neurons. RESULTS: It was shown that only lobules VIII was supplied by afferents from LCx. The FB-labeled neurons were distributed bilaterally with ipsilateral predominance in both LC and LCα, and were much more frequent in LC. They occupied the lateral region of LC and dorsolateral region of LCα. These neurons were the most numerous in the caudal part of LC, but in LCα they clustered in its rostral part. Nearly all FB-labeled neurons displayed immunoreactivity to DBH and only the single cells stained simultaneously for DBH and SOM, and for DBH and NPY. However, the FB-labeled perikarya did not contain immunoreactivities to the remaining substances investigated. Moreover, some FB- or FB/DBH-positive somata were closely apposed by SOM-, NPY- or LENK-immunoreactive fibers. CONCLUSIONS: This study has provided new data on the distribution of neurons projecting from LCx to the lobule VIII, and has revealed that they are noradrenergic (and non-cholinergic) in nature and that their activity can be modulated by some neuropeptides. The findings seem to be important considering the role of the lobule VIII in innervation of axial and proximal forelimb muscles.
14

Gender-related differences of motoreurons morphometric properties of the rat medial gastrocnemius muscle

88%
Mierzejewska-Krzyzowska B., Bukowska D., Szydlowska J., Drzymala-Celichowska H., Celichowski J.
Acta Neurobiologiae Experimentalis
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2011
|
tom 71
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nr S
Gender differences are a well-known phenomenon in animal kingdom and concern body mass and morphology. The aim of this study was to determine the gender differences in the number and size of alpha and gamma motoneurons in motor nucleus of the medial gastrocnemius (MG) in the rat. The retrogradely labeled cell bodies of motoneurons of the same age male and female Wistar rats were studied following a bath of the proximal stump of the transected MG nerve in the horseradish peroxidase solution. The number and the soma diameters of male and female MG motoneurons were determined from serial microscopic images of stained sections using the Sony CCD-Iris Camera and MultiScanBase System. It was shown that weight of the central nervous system was on average 19% higher in males than in females. The mean number of motoneurons was 11% higher in males than in females and amounted to 95 and 86 motoneurons, respectively. In each case, the average soma diameters of motoneurons in motor nucleus were distributed bimodally: motoneurons smaller than 32.5 micrometer were recognized as gamma and greater ones as alpha motoneurons. In the present material the percentage composition of gamma (22%) and alpha (78%) motoneurons were very similar in both sexes. The mean number of alpha motoneurons was 13% higher in males than in females, but differences between the mean soma diameter in the two kinds of motoneurons, gamma and alpha, in both sexes were not significant. It is concluded that a total number of motoneurons in the rat MG motor nucleus in males and females is different. Whereas in females the number of alpha and gamma motoneurons is smaller than in males, the size of motoneurons is similar. Therefore, the gender differences in weight of the central nervous system reflect rather a difference in a total number of neurons than in their size.
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