Israeli police have deployed 1,000 troops and erected roadblocks at Jerusalem's Old City ahead of the weekly Friday prayers.
Under the fresh Israeli restrictions, male worshippers under 50 were banned from performing the weekly prayers in the Al-Aqsa Mosque.
"All Muslim men under 50 and West Bankers will be barred from entering the Al-Aqsa Mosque compound for Friday prayers while women of all ages were granted access," Sheikh Azzam al-Khatib, head of the Jordan-run Organization for Muslim Endowments and Al-Aqsa Affairs, told Anadolu Agency.
Al-Khatib said that while Israel restricts the entry of Palestinians into Al-Aqsa mosque compound, it facilitates the access of Jewish settlers into the holy site.
He said that at least 1,300 Jewish settlers and 350 Israeli soldiers had forced their way into the holy site during Jewish holidays in the past ten days.
"Meantime, Israeli police banned more than 5,000 Muslims from entering the Al-Aqsa Mosque during the same period," he said.
Scores of Palestinians were injured in clashes with Israeli forces inside the Al-Aqsa Mosque compound last week.
Hamas leader decries Israeli "crimes"
The leader of Hamas on Thursday called on Muslims to defend the al-Aqsa mosque compound in Jerusalem, saying Israel was trying to seize the site.
Khaled Meshaal, speaking in the Qatari capital Doha where he lives in exile, said: "We call on all our people inside the country to hurry up to al-Aqsa to defend it."
Meshaal said the Israeli government was taking advantage of Arab preoccupation with regional turmoil, particularly in Syria and Iraq, to try to take over al-Aqsa.
"We call on the nation to be angry and to send a message of painful anger to the world that the Palestinian people, the Arab and Muslim nation, will not be silent at the Israeli crime," Meshaal said.
Asked if he was worried his call may lead to a new conflict soon after Palestinians in the Gaza Strip endured a 50-day war with Israel that killed more than 2,000 Palestinians, Meshaal said: "Nobody wants a war, but it's our right to resist and preserve our rights. We are under occupation ... We have been resisting for one hundred years and will continue.
"Al-Aqsa is worth us becoming martyrs for, and anyone who can carry a weapon in the region should go and defend it, as this is the true meaning of jihad."
In recent months, groups of extremist Jewish settlers – often accompanied by Israeli security forces – have repeatedly forced their way into East Jerusalem's flashpoint Al-Aqsa Mosque complex.
The frequent violations anger Palestinian Muslims and occasionally lead to violent confrontations.
For Muslims, Al-Aqsa represents the world's third holiest site.
Jews, for their part, refer to the area as the "Temple Mount," claiming it was the site of two prominent Jewish temples in ancient times.
Israel occupied East Jerusalem during the 1967 Middle East War. It later annexed the holy city in 1980, claiming it as the capital of the self-proclaimed Jewish state – a move never recognized by the international community.
In September 2000, a visit to the site by controversial Israeli leader Ariel Sharon sparked what later became known as the "Second Intifada" – a popular uprising against the Israeli occupation in which thousands of Palestinians were killed.
Source: World Bulletin